BUV737 Rat Anti-Human IL-2
Two color flow cytometric analysis of IL-2 expression in activated human peripheral blood lymphocytes. Human peripheral blood mononuclear cells were stimulated for 5 hours with Phorbol 12-Myristate 13-Acetate (PMA, Sigma P-8139; 50 ng/ml) and Calcium Ionophore A23187 (Sigma C-9275; 1 µg/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 and permeabilized with BD Cytofix/Cytoperm™ Fixation and Permeabilization Solution (Cat. No. 554722). The cells were then washed and stained in BD Perm/Wash™ Buffer (Cat. No. 554723) with APC Mouse Anti-Human CD3 antibody (Cat No. 555335/561810/561811) and either BD Horizon™ BUV737 Rat IgG2a, κ Isotype Control (Cat No. 612760; Left Plot) or BD Horizon BUV737 Rat Anti-Human IL-2 antibody (Cat No. 612836; Right Plot) using BD Biosciences Intracellular Cytokine Staining Protocol. The two-color flow cytometric contour plots showing correlated expression of IL-2 (or Ig Isotype control staining) versus CD3 were 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 Cell Analyzer System and FlowJo™ software.
BD Horizon™ BUV737 Rat Anti-Human IL-2
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