Recombinant:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Functional Assay:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
SDS Page:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Clear Native PAGE:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Western Blot:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Mass Spectrometry:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Labeling:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Sequencing:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Comparison:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Residue:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Construct:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Mutagenesis:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Transfection:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Molecular Weight:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Marker:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Incubation:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Two Tailed Test:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Control:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Confocal Microscopy:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Plasmid Preparation:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Affinity Purification:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Purification:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Expressing:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Knockdown:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Activity Assay:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Immunofluorescence:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
Immunoprecipitation:Article Title: Selective GRP94 inhibitors and uses thereof
Article Snippet: Protein concentrations were determined using BCA kit (Pierce) according to the manufacturer's instructions.BCA kit (Pierce) according to the manufacturer's instructions. ... The protein lysates (5-50 pg) were electrophoretically resolved by SDS-PAGE, transferred onto nitrocellulose membranes and probed with the indicated primary antibodies against: HER2 (Zymed, 28004), Hsp70 (Stressgen, SPA-810), Grp94 (Stressgen, SPA-850), Hsp90α (Abcam, Ab2928), Hsp90β (StressMarq, SMC-107B), Grp78 (Cell Signaling, 3183), Raf-1 (Santa Cruz, sc-133), phospho-Raf-1 (Cell Signaling, 9421), MEK1/2 (Cell Signaling, 8727), phospho-MEK1/2 (Cell Signaling, 9154), ERK1/2 (Cell Signaling, 4695), phospho-ERK1/2 (Cell Signaling, 4370), AKT (Cell Signaling, 9272), GM130 (Cell Signaling, 2296), Flotillin 2 (Cell Signaling, 3436), Histone H4 (Cell Signaling, 2592), Histone H1 (Santa Cruz, sc-8030), Caspase 3 (Cell Signaling, 9665), cleaved PARP (Promega, G7341), a-tubulin (Sigma, T5168) and 3-actin (Sigma, A1978).. After washing off the excess antibodies, the membranes were incubated with the corresponding horseradish peroxidase (HRP) conjugated secondary antibody.After washing off the excess antibodies, the membranes..
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), from Cell Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with app
Article Title: Phosphorylation-driven epichaperome assembly is a regulator of cellular adaptability and proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; RRID:AB_854214; 1:2000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1000), HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000) and HOP (SRA-1500; RRID:AB_10618972; 1:1000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) and anti-HA tag (ab9110, RRID:AB_307019; 1:1000) from Abcam; p-MEK1/2 (S217/221) (9154; RRID:AB_2138017; 1:1000), MEK1/2 (9122; RRID:AB_823567; 1:1000), p-mTOR (S2448) (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1000), CDC37 (4793; RRID:AB_10695539; 1:1000), HOP (5670; RRID:AB_10828378; 1:1000), p-S6 ribosomal protein (Ser235/236) (4858; RRID:AB_916156; 1:2000), S6 ribosomal protein (2217; RRID:AB_331355; 1:3000), Oct4 (2840, RRID:AB_2167691, 1:2000), p-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), CK2α (2656, RRID: AB_2236816, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 1:2000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1000) from Fisher Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-conjugated secondary antibodies: goat ant
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then 26 blocked for 1 h in 5% BSA in TBS/0.1% Tween 20. .. The blots were then probed with the following 27 antibodies: HSP90β (SMC-107; RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; 28 RRID:AB_2119373; 1:1,000) from Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), 29 and HOP (SRA-1500; RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; 30 RRID:AB_303423; 1:6,000), AHA1 (ab56721, RRID:AB_2273725, 1:1000) from Abcam; CDC37 31 (4793; RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), from Cell 32 Signaling Technologies. .. The blots were washed with TBS/0.1% Tween 20 and incubated with 33 app
Article Title: Phosphorylation-Driven Epichaperome Assembly: A Critical Regulator of Cellular Adaptability and Proliferation
Article Snippet: were then blocked for 1 h in 5% BSA in TBS/0.1% 1 Tween 20 and incubated overnight with the indicated antibodies. .. HSP90β (SMC-107; 2 RRID:AB_854214; 1:2,000) and HSP110 (SPC-195; RRID:AB_2119373; 1:1,000) from 3 Stressmarq; HSC70 (SPA-815; RRID:AB_10617277; 1:1,000), and HOP (SRA-1500; 4 RRID:AB_10618972; 1:1,000) from Enzo; HSP90α (ab2928; RRID:AB_303423; 1:6,000), AHA1 5 (ab56721, RRID:AB_2273725, 1:1,000) from Abcam; p-MEK1/2 (S217/221) (9154; 6 RRID:AB_2138017; 1:1,000), MEK1/2 (9122; RRID:AB_823567; 1:1,000), p-mTOR (S2448) 7 (5536; RRID:AB_10691552; 1:500), mTOR (2983; RRID:AB_2105622; 1:1,000), CDC37 (4793; 8 RRID:AB_10695539; 1:1,000), HOP (5670; RRID:AB_10828378; 1:1,000), p-S6 ribosomal 9 protein (Ser235/236) (4858; RRID:AB_916156; 1:2,000), S6 ribosomal protein (2217; 10 RRID:AB_331355; 1:3,000), Oct4 (2840, RRID:AB_2167691, 1:2,000), p-AKT (S473) (9271, 11 RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), HSP70 (ADI-SPA-810, 12 RRID:AB_10616513, 1:2000) from Cell Signaling Technologies, β-actin (A1978, RRID: 13 AB_476692, 1:3000) from Sigma-Aldrich, and mCherry (PA5-34974, RRID:AB_2552323, 14 1:2,000) and p-Ser226 HSP90β (PA5-105480, RRID:AB_2816908, 1:1,000) from Fisher 15 Scientific. .. The blots were washed with TBS/0.1% Tween 20 and incubated with appropriate HRP-16 conjugated secondary antibodies: goat an
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation.
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) andHSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) fromPromega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) andCdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: antibodies used in this study are listed below. .. HSP70 (ADI-SPA-810, RRID:AB_10616513, 1:2000), HSC70 (ADI-SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000), HSP60 (ADI-SPA-806, RRID:AB_10617232, 1:1000) and HSP40 (SPA-400, RRID:AB_11180881, 1:3000) were purchased from Enzo; HSP90β (SMC-107, RRID:AB_854214, 1:4000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were from Stressmarq; HSP90α (ab2928, RRID:AB_303423, 1:5000), AHA1 (ab56721, RRID:AB_2273725, 1:1000), caspase 10 (ab177475, RRID: AB_2924729, 1:1000) and Securin (ab3305, RRID:AB_2173413, 1:1000) from Abcam; cleaved PARP (G734A, RRID:AB_430876, 1:1000) from Promega; CDC37 (4793, RRID:AB_10695539, 1:3000), phospho-AKT (S473) (9271, RRID:AB_329825, 1:2000), AKT (4691, RRID:AB_915783, 1:3000), phospho-ERK (T202/Y204) (4377, RRID:AB_331775, 1:3000), ERK (4695, RRID:AB_390779, 1:5000), MCL1 (5453, RRID:AB_10694494, 1:1000), RAF1 (12552, RRID:AB_2728706, 1:1000), BCL6 (14895, RRID:AB_2798638, 1:1000), eIF4E (2067, RRID:AB_2097675, 1:2000), phospho-eIF4E (S209) (9741, RRID:AB_331677, 1:500), Eg5 (14404, RRID:AB_2798473, 1:1000), phospho-STAT3 (Y705) (9145, RRID:AB_2491009, 1:1000), STAT3 (9139, RRID:AB_331757, 1:2000), Aurora A (91590, RRID:AB_2800171, 1:1000), PLK1 (4513, RRID:AB_2167409, 1:2000), TPX2 (12245, RRID:AB_2716832, 1:250), EGFR (4267, RRID:AB_2246311, 1:1000), Cyclin A2 (67955, AB_2909603, 1:1000), NuMA (ABE1361, RRID:AB_2892052, 1:1000) from EMD Millipore; NuMA (NB100-74636, RRID:AB_1049265, 1:1000) from Novus Biologicals; Cyclin B1 (554178, RRID:AB_395289, 1:1000), Nek2 (610593, RRID:AB_397933, 1:1000) and Aurora-A (610938, RRID:AB_398251, 1:1000) were from BD Transduction Laboratories; Cyclin A (sc H-432, RRID:AB_631329, 1:1000) and Cdc20 (sc-8358, RRID:AB_2291311, 1:1000) were from Santa Cruz. .. For protein quantification, films were scanned and anal
Article Title: Systems-level analyses of protein-protein interaction network dysfunctions via epichaperomics identify cancer-specific mechanisms of stress adaptation
Article Snippet: native-cognate antibodies were used in native-PAGE analysis of epichaperome assemblies. .. HSP90β (SMC-107, RRID:AB_854214, 1:3000) and HSP110 (SPC-195, RRID:AB_2119373, 1:2000) antibodies were purchased from Stressmarq; HSP70 (SPA-810, RRID:AB_10616513, 1:2000), HSC70 (SPA-815, RRID:AB_10617277, 1:3000), HOP (SRA-1500, RRID:AB_10618972, 1:2000) from Enzo; HSP90α (ab2928, RRID:AB_303423, 1:6000) from Abcam.
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