Images for product: anti-Tumor Necrosis Factor (TNF) antibody
Expression of TNF by stimulated CD4+ and CD4- BALB/c spleen cells. After 6 hour stimulation with hamster anti-mouse CD3 (2 µg/ml final concentration, Clone 145-2C11) and hamster anti-mouse CD28 (2 µg/ml final concentration, Clone 37.51) antibodies in the presence of GolgiStop™ (2 µM final concentration), the splenocytes were stained with FcBlock™ (1 µg/1 million cells), then 0.06 µg of PE-conjugated rat anti-mouse CD4 (PE-RM4-5). The cells were then fixed, permeabilized, and subsequently stained with 0.06 µg of FITC-conjugated rat anti-mouse TNF antibody (FITC-MP6-XT22) by using Pharmingen's staining protocol (first panel). To demonstrate specificity of staining, the binding of the FITC-MP6-XT22 antibody was blocked by preincubation of the antibody conjugate with recombinant mouse TNF (0.25 µg, middle panel), and by preincubation of the fixed/permeabilized cells with unlabeled MP6-XT22 antibody (2 µg, ABIN967462, second panel) prior to staining. The quadrant markers for the bivariate dot plots were set based on the autofluorescence control, and verified with the recombinant cytokine blocking (middle panel) and antibody blocking (second panel) specificity controls.