BD HorizonTM R718 Mouse Anti-Human IL-2

Description

Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software. Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software. Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software. Show More Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software. Show More Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software. Two color flow cytometric analysis of IL-2 expression by activated Human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 h with Phorbol 12-Myristate 13-Acetate PMA, Sigma P-8139; 50 ng/ml and Calcium Ionophore A23187 Sigma C-9275; 1 mu g/ml , in the presence of BD GolgiStop Protein Transport Inhibitor containing Monensin Cat. No. 554724 . The cells were harvested, washed with BD Pharmingen Stain Buffer FBS Cat. No. 554656 and fixed with BD Cytofix Fixation Buffer Cat. No. 554655 followed by washing and permeabilization using BD Perm/Wash Buffer Cat. No. 554723 . The permeabilized cells were then stained in BD Perm/Wash Buffer with PE Mouse Anti-Human CD3 antibody Cat. No. 555333 and with either BD Horizon R718 Mouse IgG1, kappa Isotype Control Cat No. 566928; Left Plot or BD Horizon R718 Mouse Anti-Human IL-2 antibody Cat No. 569490/569491; Right Plot . The pseudocolor density plot showing the correlated expression of IL-2 or Ig Isotype control staining versus CD3 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 LSRFortessa X-20 Flow Cytometer System and FlowJo software.