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Two-color flow cytometric analysis of IFN-γ expression in stimulated human peripheral blood mononuclear cells. HiCK-1 Human Cytokine Positive Control Cells (Cat. No. 555061) were permeabilized with BD Perm/Wash™ Buffer (Cat. No. 554723). The cells were then stained with APC Mouse Anti-Human CD3 antibody (Cat. No. 561811/555335/561810) and either BD Horizon™ BUV395 Mouse IgG1, κ Isotype Control (Cat No. 563547, Left Panel) or BD Horizon™ BUV395 Mouse Anti-Human IFN-γ antibody (Cat No. 563563/565971, Right Panel). Two-color flow cytometric dot plots showing the expression of IFN-γ (or Ig Isotype control staining) versus CD3 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.