BD PhosflowTM PE Mouse anti-Src (pY418)

Description

Analysis of Src pY418 in human T leukemia cells and peripheral blood lymphocytes. Jurkat cells ATCC TIB152 , left panel and human peripheral blood mononuclear cells PBMC, middle panel were either treated with 1 mu M Staurosporine EMD Biosciences, Cat. No. 569397 for 2 hours 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 with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Src pY418, Cat. No. 560094 . Lymphocytes were selected by their scatter profile during analysis of the PBMC. The data demonstrates that the level of phosphorylation of Src pY418 decreases when protein kinase activity is inhibited by the treatment. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. The specificity of mAb K98-37 was confirmed by western blot analysis right panel using unconjugated Rabbit anti-Src Cell Signaling Technology, Cat. No. 2123, left blot and unconjugated Mouse anti-Src pY418 right blot monoclonal antibodies on lysates of Jurkat cells that were untreated lanes 1 or treated with Staurosporine for 1 hour lanes 2 or 2 hours lanes 3 . Src pY418 is identified as a band of 60 kDa that is reduced by Staurosporine treatment. Analysis of Src pY418 in human T leukemia cells and peripheral blood lymphocytes. Jurkat cells ATCC TIB152 , left panel and human peripheral blood mononuclear cells PBMC, middle panel were either treated with 1 mu M Staurosporine EMD Biosciences, Cat. No. 569397 for 2 hours 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 with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Src pY418, Cat. No. 560094 . Lymphocytes were selected by their scatter profile during analysis of the PBMC. The data demonstrates that the level of phosphorylation of Src pY418 decreases when protein kinase activity is inhibited by the treatment. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. The specificity of mAb K98-37 was confirmed by western blot analysis right panel using unconjugated Rabbit anti-Src Cell Signaling Technology, Cat. No. 2123, left blot and unconjugated Mouse anti-Src pY418 right blot monoclonal antibodies on lysates of Jurkat cells that were untreated lanes 1 or treated with Staurosporine for 1 hour lanes 2 or 2 hours lanes 3 . Src pY418 is identified as a band of 60 kDa that is reduced by Staurosporine treatment. Show More Analysis of Src pY418 in human T leukemia cells and peripheral blood lymphocytes. Jurkat cells ATCC TIB152 , left panel and human peripheral blood mononuclear cells PBMC, middle panel were either treated with 1 mu M Staurosporine EMD Biosciences, Cat. No. 569397 for 2 hours 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 with BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes, and then stained with PE Mouse anti-Src pY418, Cat. No. 560094 . Lymphocytes were selected by their scatter profile during analysis of the PBMC. The data demonstrates that the level of phosphorylation of Src pY418 decreases when protein kinase activity is inhibited by the treatment. Flow cytometry was performed on a BD FACSCalibur flow cytometry system. The specificity of mAb K98-37 was confirmed by western blot analysis right panel using unconjugated Rabbit anti-Src Cell Signaling Technology, Cat. No. 2123, left blot and unconjugated Mouse anti-Src pY418 right blot monoclonal antibodies on lysates of Jurkat cells that were untreated lanes 1 or treated with Staurosporine for 1 hour lanes 2 or 2 hours lanes 3 . Src pY418 is identified as a band of 60 kDa that is reduced by Staurosporine treatment.