Apoptosis Assay:Article Title: ANP32E contributes to gastric cancer progression via NUF2 upregulation.
Article Snippet: After transfection for 48 h at 37 ̊C, the cells were resuspended using 1X annexin‐binding buffer and were stained using an alexa Fluor 488, annexin V and Pi cell apoptosis analysis kit (Thermo Fisher Scientific, Inc.), according to the manu‐ facture's protocols. .. PAC‐1 (2 μM; Selleck Chemicals; cat. no. S2738) was used to induce cell apoptosis, then the cells were subjected to cell apoptosis assay. .. Subsequently, a cyto‐ FLEX flow cytometer (Beckman Coulter, Inc.) was used to detect cell apoptosis. ur plus lr quadrants were used to calculate apoptosis.
Article Title: ANP32E contributes to gastric cancer progression via NUF2 upregulation
Article Snippet: After transfection for 48 h at 37°C, the cells were resuspended using 1X annexin-binding buffer and were stained using an Alexa Fluor 488, annexin V and PI cell apoptosis analysis kit (Thermo Fisher Scientific, Inc.), according to the manufacture's protocols. .. PAC-1 (2 μM; Selleck Chemicals; cat. no. S2738) was used to induce cell apoptosis, then the cells were subjected to cell apoptosis assay. .. Subsequently, a cytoFLEX flow cytometer (Beckman Coulter, Inc.) was used to detect cell apoptosis.
TUNEL Assay:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
Activity Assay:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
Purification:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
cDNA Synthesis:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
Recombinant:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
In Situ:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
Binding Assay:Article Title: Microbes exploit death-induced nutrient release by gut epithelial cells.
Article Snippet: .. The reagents used for different parts of this work were obtained from the indicated suppliers as follows: doxycycline (Sigma D-9891); B/B homodimerizer (Clonetech AP20187); QVD (Sigma SML0063); staurosporine (Abcam ab120056); TUNEL (Sigma 12156792910); CD45 antibody (Abcam ab10558); caspase 3 activity kit (Sigma APT131, AssayGenie RG BN00018); caspase 8 activity kit (Sigma APT129); annexin V-APC (Biolegend 640941); annexin V-Pac Blue (Biolegene 640917); TO-PRO-3 Iodide (Thermo Fisher T3605); 7AAD (Invitrogen A1310); Yoyo-1 (Thermo Fisher Y3601); Sytox blue (Thermo Fisher S34857); PAC-1 (Selleckchem S2738); doxorubicin (Sigma D-1515); proteinase K (GC Biotech BIO-37037); RiboPure RNA Purification Kit (Thermo Fisher AM1925); SensiFast cDNA Synthesis Kit (GC Biotech BIO-650504); recombinant human TNF (VIB Protein Core); inosine monophosphate (IMP; Sigma 57510); dihydroxyacetone phosphate (DHAP; Sigma 37442); guanosine monophosphate (GMP; Sigma G8377); UDP-glucose (Abcam ab120384); spermidine (Sigma S2626); FBP (Sigma F6803); In Situ Cell Death Detection Kit, TUNEL (Sigma 12156792910); annexin V binding buffer (BD 556454); shikonin (MedChemExpress HY-N0822); pyruvate detection kit (Merck MAK071); formate detection kit (Sigma MAK059); FBP detection kit (Biovision K2036). .. The following antibodies were used: caspase-1 (rabbit, anti-mouse)41, caspase-3 (CST 9662), cleaved caspase-3 (CST 9664), caspase-7 (CST 8438), cleaved caspase-7 (Abcam ab255818), caspase-8 (Abnova MAB3429), cleaved caspase-8 (CST 9429), caspase-8 (CST 9746), P-MLKL Ser345 (Abcam ab196436), MLKL (Sigma-Aldrich MABC604), P-RIPK3 Thr231/232 (CST 91702), tubulin HRP (Abcam ab21058), sheep anti-mouse IgG HRP (Cytiva NA931), goat anti-rat IgG HRP (Cytiva NA935), goat anti-rabbit IgG HRP (Cayman Chemical 10004301).
other:
Titration:Article Title: A clinical-stage oncology compound selectively targets drug-resistant cancers
Article Snippet: Cells were seeded at 2000 cells/well into 12-well Cell+ plates (Sarstedt). .. The following day a titration of PAC-1 (1, 2.5, 5, 7.5, 10, 15, 20, 25 μM, SelleckChem) was added and cells incubated with the drug continually for 21 days, with the media changed every 5–7 days. .. Plates were fixed in 1% formaldehyde, before being stained with a 0.05% v/v crystal violet (Sigma Aldrich) solution.
Incubation:Article Title: A clinical-stage oncology compound selectively targets drug-resistant cancers
Article Snippet: Cells were seeded at 2000 cells/well into 12-well Cell+ plates (Sarstedt). .. The following day a titration of PAC-1 (1, 2.5, 5, 7.5, 10, 15, 20, 25 μM, SelleckChem) was added and cells incubated with the drug continually for 21 days, with the media changed every 5–7 days. .. Plates were fixed in 1% formaldehyde, before being stained with a 0.05% v/v crystal violet (Sigma Aldrich) solution.
|
Selleck Chemicals is a verified supplier 
Selleck Chemicals manufactures this product 
