PE-Cy™7 Mouse IgG1 κ Isotype Control
Ig isotype control for the expression of IFN-γ by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated for 4 hours with PMA (5 ng/ml, Sigma) and Ionomycin (500 ng, Sigma) in the presence of Brefeldin A (GolgiPlug, Cat. No. 555029). Cells were harvested, fixed, permeabilized and stained with PE Mouse Anti-Human CD8 (Cat. No. 555367) and either PE-Cy™7 Mouse Anti-Human IFN-γ (Cat. No. 557643; left panel) or PE-Cy™7 Mouse IgG1 κ Isotype Control (Cat. No. 557646/565573/557872; right panel) as a specificity control by using Pharmingen's staining protocol. To demonstrate additional specificity of staining the binding of PE-Cy™7 Mouse Anti-Human IFN-γ was blocked by preincubation of the fixed/permeabilized cells with an excess of unlabeled Purified Mouse Anti-Human IFN-γ (5 µg, Cat. No. 554699; data not shown) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence and isotype control.
BD Pharmingen™ PE-Cy™7 Mouse IgG1 κ Isotype Control
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