PE Mouse IgG1, κ Isotype Control
Expression of IFN-γ by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma, Cat. No. P-8139) and calcium ionophore A23187 (Sigma, Cat. No. C-9275), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy5 anti-CD3 (PE-Cy5-UCHT1, Cat. No. 555334), fixed, permeabilized, and subsequently stained with 0.25 µg of PE-mouse anti-human IFN-γ antibody (PE-4S.B3, Cat. No. 554552, left panel) or with 0.25 µg PE-MOPC-21 immunoglobulin (Cat. No. 554680, right panel) using the BD Pharmingen™ staining protocol. To demonstrate specificity of staining, the binding of PE-4S.B3 antibody was blocked by preincubation of fixed/permeabilized cells with excess unlabelled 4S.B3 antibody (5 µg; Cat. No. 554549). The quadrant markers for the bivariate dot plot were set based on autofluorescence controls and verified using the unlabelled 4S.B3 antibody blocking control.
BD Pharmingen™ PE Mouse IgG1, κ Isotype Control
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