BD PhosflowTM Alexa Fluor 488 Anti-Stat5 (pY694)

Description

Flow cytometric analysis of phospho-Stat5 pY694 . TF-1 cells GM-CSF dependent cell line were starved overnight in IMDM with 0.1 % FBS without GM-CSF. The following day, cells were either left unstimulated unshaded or stimulated with GM-CSF recombinant protein Cat. No. 550068 at 25 ng/ml for 15 minutes at 37C shaded . Cells were fixed using BD Cytofix Fixation Buffer 10 minutes at 37C and then permeabilized in BD Phosflow Perm Buffer III 30 minutes on ice or overnight at -20C . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with Alexa Fluor 488-conjugated phospho-Stat5 pY694 antibody 20 mul/1x10e6 cells for 1 hour at RT; clone 47 . The cells were analyzed on a BD FACSCalibur flow cytometer. Flow cytometric analysis of phospho-Stat5 pY694 . TF-1 cells GM-CSF dependent cell line were starved overnight in IMDM with 0.1 % FBS without GM-CSF. The following day, cells were either left unstimulated unshaded or stimulated with GM-CSF recombinant protein Cat. No. 550068 at 25 ng/ml for 15 minutes at 37C shaded . Cells were fixed using BD Cytofix Fixation Buffer 10 minutes at 37C and then permeabilized in BD Phosflow Perm Buffer III 30 minutes on ice or overnight at -20C . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with Alexa Fluor 488-conjugated phospho-Stat5 pY694 antibody 20 mul/1x10e6 cells for 1 hour at RT; clone 47 . The cells were analyzed on a BD FACSCalibur flow cytometer. Show More Flow cytometric analysis of phospho-Stat5 pY694 . TF-1 cells GM-CSF dependent cell line were starved overnight in IMDM with 0.1 % FBS without GM-CSF. The following day, cells were either left unstimulated unshaded or stimulated with GM-CSF recombinant protein Cat. No. 550068 at 25 ng/ml for 15 minutes at 37C shaded . Cells were fixed using BD Cytofix Fixation Buffer 10 minutes at 37C and then permeabilized in BD Phosflow Perm Buffer III 30 minutes on ice or overnight at -20C . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with Alexa Fluor 488-conjugated phospho-Stat5 pY694 antibody 20 mul/1x10e6 cells for 1 hour at RT; clone 47 . The cells were analyzed on a BD FACSCalibur flow cytometer.