Images for product: anti-Interleukin 4 (IL4) antibody
Expression of IL-4 by stimulated human peripheral blood mononuclear cells (PBMC). Human PBMC were stimulated with soluble anti-human CD3 antibody (1 µg/ml), recombinant human IL-2 (10 ng/ml) and IL-4 (20 ng/ml) for 2 days. The cells were subsequently cultured in medium containing recombinant human IL-2 and recombinant human IL-4 for 3 days. Finally, the cells were harvested and stimulated for 6 h with PMA (Sigma) and calcium ionophore A23187 (Sigma) in the presence of 2 µM GolgiStop™. The cells were harvested, stained with PE-Cy™5 -anti CD4, fixed, permeabilized, and subsequently stained with 0.05 µg of PE-rat anti-human IL-4 antibody (PE-MP4-25D2) by using Pharmingen's staining protocol (first panel). To demonstrate specificity of staining, the binding of PE-MP4-25D2 antibody was blocked by preincubation of the fixed/permeabilized cells with unlabeled MP4-25D2 antibody (2.5 µg, ABIN967463, second panel) prior to staining. The quadrant markers for the bivariate dot plot were set based on the autofluorescence controls and verified using unlabeled antibody blocking controls.