BD PhosflowTM Alexa Fluor 647 Mouse Anti-mTOR (pS2448)

Description

Two-color flow cytometric analysis of mTOR pS2448 expression in stimulated human peripheral blood B cells. Human peripheral blood mononuclear cells PBMC were either not stimulated dashed line histogram or stimulated solid line histogram with Type C CpG oligonucleotide 1 muM, 4 hrs; InvivoGen, Cat. No. TLRL-2395 . The cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655; 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III Cat. No. 558050; 30 minutes on ice . The cells were washed twice with BD Pharmingen Stain Buffer Cat. No. 554656 and then stained with BD Phosflow Alexa Fluor 647 Mouse Anti-mTOR pS2448 Cat. No. 564242 and BD Phosflow PE Mouse Anti-Human CD20 Cat. No. 561174 antibodies using the BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from CD20 positive-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Two-color flow cytometric analysis of mTOR pS2448 expression in stimulated human peripheral blood B cells. Human peripheral blood mononuclear cells PBMC were either not stimulated dashed line histogram or stimulated solid line histogram with Type C CpG oligonucleotide 1 muM, 4 hrs; InvivoGen, Cat. No. TLRL-2395 . The cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655; 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III Cat. No. 558050; 30 minutes on ice . The cells were washed twice with BD Pharmingen Stain Buffer Cat. No. 554656 and then stained with BD Phosflow Alexa Fluor 647 Mouse Anti-mTOR pS2448 Cat. No. 564242 and BD Phosflow PE Mouse Anti-Human CD20 Cat. No. 561174 antibodies using the BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from CD20 positive-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System. Show More Two-color flow cytometric analysis of mTOR pS2448 expression in stimulated human peripheral blood B cells. Human peripheral blood mononuclear cells PBMC were either not stimulated dashed line histogram or stimulated solid line histogram with Type C CpG oligonucleotide 1 muM, 4 hrs; InvivoGen, Cat. No. TLRL-2395 . The cells were fixed with BD Cytofix Fixation Buffer Cat. No. 554655; 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III Cat. No. 558050; 30 minutes on ice . The cells were washed twice with BD Pharmingen Stain Buffer Cat. No. 554656 and then stained with BD Phosflow Alexa Fluor 647 Mouse Anti-mTOR pS2448 Cat. No. 564242 and BD Phosflow PE Mouse Anti-Human CD20 Cat. No. 561174 antibodies using the BD Biosciences Protocol for Intracellular Staining. The fluorescence histograms were derived from CD20 positive-gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD LSRFortessa Cell Analyzer System.