BD PhosflowTM Alexa Fluor 647 Mouse Anti-Stat5 (pY694)

Description

Flow cytometric analysis of Stat5 pY694 . TF-1 cells Human erythroleukemia; ATCC CRL-2003 , a 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 shaded with 25 ng/ml GM-CSF recombinant protein Cat. No. 550068 for 15 minutes at 37C. Cells were fixed with BD Cytofix buffer 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III 30 minutes on ice . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with the Alexa Fluor 647 mouse anti-Stat5 pY694 antibody. The cells were analyzed on a BD FACSCalibur flow cytometer. For intracellular staining of human whole blood, BD Phosflow Lyse/Fix buffer MN 558049 may be used for fixation. Flow cytometric analysis of Stat5 pY694 . TF-1 cells Human erythroleukemia; ATCC CRL-2003 , a 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 shaded with 25 ng/ml GM-CSF recombinant protein Cat. No. 550068 for 15 minutes at 37C. Cells were fixed with BD Cytofix buffer 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III 30 minutes on ice . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with the Alexa Fluor 647 mouse anti-Stat5 pY694 antibody. The cells were analyzed on a BD FACSCalibur flow cytometer. For intracellular staining of human whole blood, BD Phosflow Lyse/Fix buffer MN 558049 may be used for fixation. Show More Flow cytometric analysis of Stat5 pY694 . TF-1 cells Human erythroleukemia; ATCC CRL-2003 , a 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 shaded with 25 ng/ml GM-CSF recombinant protein Cat. No. 550068 for 15 minutes at 37C. Cells were fixed with BD Cytofix buffer 10 minutes at 37C and then permeabilized by adding BD Phosflow Perm Buffer III 30 minutes on ice . Cells were then washed twice in BD Pharmingen Stain Buffer Cat. No. 554656 , and stained with the Alexa Fluor 647 mouse anti-Stat5 pY694 antibody. The cells were analyzed on a BD FACSCalibur flow cytometer. For intracellular staining of human whole blood, BD Phosflow Lyse/Fix buffer MN 558049 may be used for fixation.