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FITC 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 ionophone A23187 (250 ng/ml final concentration; Sigma, Cat. No. C-9275) in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PBMC were stained with PE-Cy5 (formerly referred to as BD-Cy-Chrome™) - anti-CD3 (Cy-Chrome™ - UCHT1, Cat. No. 555334), fixed, permeabilized, and subsequently stained with 0.25 µg of FITC-mouse anti-human IFN-γ antibody (FITC-4S.B3, Cat. No. 554551; left panel) or FITC-MOPC-21 immunoglobulin (0.25 µg; Cat. No. 554679; right panel) by using the BD Bioscience staining protocol. To demonstrate specificity of staining, the binding of FITC-4S.B3 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™ FITC Mouse IgG1, κ Isotype Control
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