APC Mouse IgG1 κ Isotype Control
Expression of TNF by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated for 6 hours with PMA (50 ng/ml final concentration; Sigma) and calcium ionophore A23187 (1 µg/ml final concentration; Sigma), in the presence of GolgiStop™ (2 µM final concentration; Cat. No. 554724). The PMBC were stained with FITC-anti-CD3 (FITC-UCHT1, Cat. No. 555332), fixed, permeabilized, and subsequently stained with 0.125 µg of APC-mouse anti-human TNF (APC-MAb11, Cat. No. 554514, left panel) or with 0.125 µg APC-mouse IgG1 (Cat. No. 554681, right panel) using BD Pharmingen™ staining protocol. To demonstrate specificity, the binding of APC-MAb11 antibody was blocked by perincubation of fixed/permabilized cells with excess unlabelled MAb11 antibody (5 µg; Cat. No. 554510). The quadrant markers for the bivariate dot plot were set based on isotype controls and verified using the unlabelled MAb11 antibody blocking control. The APC-conjugated reagent can be used in any flow cytometer equipped with a dye, HeNE or red diode laser. These include the dual laser FACStarPLUS™, FACSVantage™, or FACSCalibur™.
BD Pharmingen™ APC Mouse IgG1 κ Isotype Control
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