BD PhosflowTM PE Mouse anti-Stat1 (pS727)

Description

Analysis of Stat1 pS727 in human epithelioid carcinoma and peripheral blood lymphocytes. HeLa S3 cells ATCC CCL 2.2, left panel were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed-line histogram or untreated solid-line histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, then permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, and then stained with PE Mouse anti-Stat1 pS727, Cat. No. 560069 . Human peripheral blood mononuclear cells PBMC, right panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The PBMC were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Stat1 pS727 . For data analysis, lymphocytes were selected by their scatter profile. All flow cytometry was performed on a BD FACSCalibur flow cytometry system. The specificity of mAb K51-856 was confirmed by western blot. Lysates from PMA-treated cells lane 1, left blot , control PBMC lane 2, left blot , Nocadazole-treated cells lane 1, right blot , and control HeLa S3 cells lane 2, right blot were treated with unconjugated antibody at 2.0 ug/ml. 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 ATCC CCL 2.2, left panel were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed-line histogram or untreated solid-line histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, then permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, and then stained with PE Mouse anti-Stat1 pS727, Cat. No. 560069 . Human peripheral blood mononuclear cells PBMC, right panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The PBMC were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Stat1 pS727 . For data analysis, lymphocytes were selected by their scatter profile. All flow cytometry was performed on a BD FACSCalibur flow cytometry system. Show More The specificity of mAb K51-856 was confirmed by western blot. Lysates from PMA-treated cells lane 1, left blot , control PBMC lane 2, left blot , Nocadazole-treated cells lane 1, right blot , and control HeLa S3 cells lane 2, right blot were treated with unconjugated antibody at 2.0 ug/ml. 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 ATCC CCL 2.2, left panel were either treated with 0.5 mug/ml Nocodazole Sigma-Aldrich, Cat. No. M1404 at 37 C for 18 hours dashed-line histogram or untreated solid-line histogram . The cells were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, then permeabilized BD Phosflow Perm Buffer III, Cat. No. 558050 on ice for at least 30 minutes, and then stained with PE Mouse anti-Stat1 pS727, Cat. No. 560069 . Human peripheral blood mononuclear cells PBMC, right panel were either stimulated with 50 nM PMA Sigma, P8139 for 15 minutes at 37oC shaded histogram or untreated open histogram . The PBMC were fixed BD Cytofix buffer, Cat. No. 554655 for 10 minutes at 37 C, permeabilized with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Stat1 pS727 . For data analysis, lymphocytes were selected by their scatter profile. All flow cytometry was performed on a BD FACSCalibur flow cytometry system. The specificity of mAb K51-856 was confirmed by western blot. Lysates from PMA-treated cells lane 1, left blot , control PBMC lane 2, left blot , Nocadazole-treated cells lane 1, right blot , and control HeLa S3 cells lane 2, right blot were treated with unconjugated antibody at 2.0 ug/ml. Stat1 pS727 is identified as a band of 91 kDa in the treated cells.