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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 μ g/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were fixed with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722), washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), and then stained with FITC Mouse Anti-Human CD3 antibody (Cat No. 555332/561806/561807) and either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat No. 562652, Left Panel) or BD Horizon™ BV605 Mouse Anti-Human IL-2 antibody (Cat No. 563947, Right Panel) by using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric dot plots show the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 and were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.
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 μ g/ml), in the presence of BD GolgiStop™ Protein Transport Inhibitor (containing Monensin) (Cat. No. 554724). The cells were fixed with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722), washed and permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723), and then stained with FITC Mouse Anti-Human CD3 antibody (Cat No. 555332/561806/561807) and either BD Horizon™ BV605 Mouse IgG1, κ Isotype Control (Cat No. 562652, Left Panel) or BD Horizon™ BV605 Mouse Anti-Human IL-2 antibody (Cat No. 563947, Right Panel) by using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric dot plots show the correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 and were derived from gated events with the forward and side light-scatter characteristics of intact lymphocytes. Flow cytometric analysis was performed using a BD™ LSR II Flow Cytometer System.