BD PhosflowTM Alexa Fluor 647 Mouse anti-Stat1 (pS727)

Description

Analysis of Stat1 pS727 in human epithelioid carcinoma and peripheral blood lymphocytes. HeLa S3 cells left panel, ATCC CCL 2.2 were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed line or untreated solid line . Human peripheral blood mononuclear cells middle panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, and then stained with Alexa Fluor 647 Mouse anti-Stat1 pS727, Cat. No. 560190 . Flow cytometry was performed on a BD LSR II left or FACSCalibur middle flow cytometer, and lymphocytes were identified by their scatter profile. The right panel shows confirmation of the specificity by western blot using unconjugated K51-856 antibody at 2.0 mug/ml on lysates from PMA-treated lane 1, left blot and control lane 2 PBMCs and Nocodazole-treated lane 1, right blot and control lane 2 HeLa S3 cells. Stat1 pS727 is identified as a band of 91 kDa in the treated cells. Analysis of Stat1 pS727 in human epithelioid carcinoma and peripheral blood lymphocytes. HeLa S3 cells left panel, ATCC CCL 2.2 were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed line or untreated solid line . Human peripheral blood mononuclear cells middle panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, and then stained with Alexa Fluor 647 Mouse anti-Stat1 pS727, Cat. No. 560190 . Flow cytometry was performed on a BD LSR II left or FACSCalibur middle flow cytometer, and lymphocytes were identified by their scatter profile. The right panel shows confirmation of the specificity by western blot using unconjugated K51-856 antibody at 2.0 mug/ml on lysates from PMA-treated lane 1, left blot and control lane 2 PBMCs and Nocodazole-treated lane 1, right blot and control lane 2 HeLa S3 cells. Stat1 pS727 is identified as a band of 91 kDa in the treated cells. Show More Analysis of Stat1 pS727 in human epithelioid carcinoma and peripheral blood lymphocytes. HeLa S3 cells left panel, ATCC CCL 2.2 were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed line or untreated solid line . Human peripheral blood mononuclear cells middle panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for 30 minutes, and then stained with Alexa Fluor 647 Mouse anti-Stat1 pS727, Cat. No. 560190 . Flow cytometry was performed on a BD LSR II left or FACSCalibur middle flow cytometer, and lymphocytes were identified by their scatter profile. The right panel shows confirmation of the specificity by western blot using unconjugated K51-856 antibody at 2.0 mug/ml on lysates from PMA-treated lane 1, left blot and control lane 2 PBMCs and Nocodazole-treated lane 1, right blot and control lane 2 HeLa S3 cells. Stat1 pS727 is identified as a band of 91 kDa in the treated cells.