BD PhosflowTM RY703 Mouse Anti-Stat5 (pY694)

Description

Flow cytometric analysis of Stat5 PY694 expression in IL-2-stimulated Human lymphocytes. Human whole blood was either left Unstimulated dashed line histogram or Stimulated solid line histogram with 100 ng/mL recombinant Human IL-2 protein Cat. No. 554603 for 15 minutes at 37C. Erythrocytes were lysed and the leukocytes were fixed in a single step using BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10 min at 37C. Cells were then washed, pelleted by centrifugation, and resuspended and permeabilized in BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes. The permeabilized cells were washed twice with 5-10 mL of BD Pharmingen Stain Buffer FBS Cat. No. 554656 and then stained with either BD Horizon RY703 Mouse IgG1, kappa Isotype Control Cat. No. 571453; dashed line histogram or BD Phosflow RY703 Mouse Anti-Stat5 pY694 antibody Cat. No. 571798/571849; solid line histogram . The histogram showing Stat5 pY694 expression or Ig Isotype control staining was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony A5 SE Cell Analyzer System and FlowJo Software. Flow cytometric analysis of Stat5 PY694 expression in IL-2-stimulated Human lymphocytes. Human whole blood was either left Unstimulated dashed line histogram or Stimulated solid line histogram with 100 ng/mL recombinant Human IL-2 protein Cat. No. 554603 for 15 minutes at 37C. Erythrocytes were lysed and the leukocytes were fixed in a single step using BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10 min at 37C. Cells were then washed, pelleted by centrifugation, and resuspended and permeabilized in BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes. The permeabilized cells were washed twice with 5-10 mL of BD Pharmingen Stain Buffer FBS Cat. No. 554656 and then stained with either BD Horizon RY703 Mouse IgG1, kappa Isotype Control Cat. No. 571453; dashed line histogram or BD Phosflow RY703 Mouse Anti-Stat5 pY694 antibody Cat. No. 571798/571849; solid line histogram . The histogram showing Stat5 pY694 expression or Ig Isotype control staining was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony A5 SE Cell Analyzer System and FlowJo Software. Show More Flow cytometric analysis of Stat5 PY694 expression in IL-2-stimulated Human lymphocytes. Human whole blood was either left Unstimulated dashed line histogram or Stimulated solid line histogram with 100 ng/mL recombinant Human IL-2 protein Cat. No. 554603 for 15 minutes at 37C. Erythrocytes were lysed and the leukocytes were fixed in a single step using BD Phosflow Lyse/Fix Buffer Cat. No. 558049 for 10 min at 37C. Cells were then washed, pelleted by centrifugation, and resuspended and permeabilized in BD Phosflow Perm Buffer III Cat. No. 558050 on ice for 30 minutes. The permeabilized cells were washed twice with 5-10 mL of BD Pharmingen Stain Buffer FBS Cat. No. 554656 and then stained with either BD Horizon RY703 Mouse IgG1, kappa Isotype Control Cat. No. 571453; dashed line histogram or BD Phosflow RY703 Mouse Anti-Stat5 pY694 antibody Cat. No. 571798/571849; solid line histogram . The histogram showing Stat5 pY694 expression or Ig Isotype control staining was derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometry and data analysis were performed using a BD FACSymphony A5 SE Cell Analyzer System and FlowJo Software.