Protein Enrichment:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Recombinant:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Luciferase:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Electron Microscopy:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Bicinchoninic Acid Protein Assay:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Proximity Ligation Assay:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Software:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Cloning:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
Purification:Article Title: Identification of Cellular Protein Targets of a Half-Sandwich Iridium(III) Complex Reveals Its Dual Mechanism of Action via Both Electrophilic and Oxidative Stresses.
Article Snippet: Antibodies Source Identifier Rabbit anti-Nrf2 Abcam #137550 Mouse anti-β-Actin Cell Signaling #4967 Mouse anti-phospho-Histone H2A.X (Ser139) Upstate technology #05-636 clone JBW301 Mouse anti-biotin Sigma #B7653 Rabbit anti-FLNB [N1] GeneTex #GTX101206 Rabbit anti-P34-Arc/ARPC2 Merck #07-227-I Rabbit anti-HSP90 Proteintech #13171-1-AP Rabbit anti-GRP78/BIP Proteintech #11587-1-AP Rabbit anti-TOMM20 Abcam #AB186735 Streptavidin-HRP conjugate R&D systems #P237707 HRP-conjugated anti-mouse IgG Cell Signaling #7076 HRP-conjugated anti-rabbit IgG Cell Signaling #7074 Alexa Fluor® 488 Goat-anti Mouse Invitrogen #A11001 Alexa Fluor® 488 Goat-anti Rabbit Invitrogen #A11008 Chemicals and proteins Source Identifier THPTA Merck #762342 Click-&-Go® protein enrichment Kit Click chemistry tools #1033 HaltTM protease and phosphatase inhibitor cocktail Thermo Fisher #78440 Trypsin/LysC Promega #V5073 Hoechst 33342 Thermo Fisher #H3570 Biotin-PEG4-alkyne Lumiprobe #2615 QuantiLum recombinant luciferase Promega #E1701 Radicicol Merck #R2146 FAM alkyne, 5-isomer Lumiprobe #C41B0 Alexa FluorTM 633 phalloidin Thermo Fisher #A22284 Poly-L-lysine (PLL) 0.1% solution in H2O Merck #P8920 PolyEthylene Glycol-Succinimidyl Valerate (PEG-SVA) Alvéole N/A Fibronectin Invitrogen #33010018 Alexa FluorTM 633 phalloidin Thermo Fisher #A22281 Glutaraldehyde 25% solution in H2O Merck #G5882 DRAQ5TM Thermo Fisher #62254 Tannic acid Merck #403040 Uranyl acetate Electron microscopy sciences #2240920 Hexamethyldisilazane Merck #379212 SPY650-FastAct Spirochrome #CS505 MitoTrackerTM Green FM Invitrogen #M7514 7-AAD (7-Aminoactinomycin D) Sigma #A9400 Mito-tracker red CMX-Ros Thermo Fisher #M46752 2-DG (2-deoxy-glucose) TCI #D0051 CCCP ((Carbonyl cyanide m-chlorophenyl hydrazone) Sigma #C2759 S4 Commercial assays Source Identifier alamarBlueTM cell viability Thermo Fisher #DAL1100 BCA protein assay Thermo Fisher #23229 DuoLink® proximity ligation assay Merck #DUO92008 Cell Lines Source Identifier HeLa-HyPer DOI: 10.1089/ars.2015.6586 N/A HeLa ATCC CCL-2 HEK293 ATCC CRL-1573 hTERT-RPE1 ATCC CRL-4000 Huh-7 PMID: 6286115 N/A Software and Algorithms Source Identifier Prism (V 5c) GraphPad N/A Protein discoverer V 2.4 Thermo Fisher link myProMS V 3.9.3 DOI: 10.1002/pmic.200600784 link MassChroQ V 2.2.21 DOI: 10.1002/pmic.201100120 N/A Percolator Biotools DOI: 10.1007/s13361-016-1460-7 (link) Leonardo Alveole link Fiji N/A link ImageLab (V 4.0.1) Biorad N/A PyMca DOI: 10.1016/j.sab.2006.12.002 link Kaluza Beckman N/A S5 Heat Shock Proteins cloning and purification Plasmids.
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