Western blot analysis of extracts from purified CD4 + human peripheral blood mononuclear cells, activated with anti-CD3 and anti-CD28 in Th2-inducing conditions for 12 d, untreated (-) or treated (+) with TPA #4174 (40 nM, 6 hr), Ionomycin #9995 (3 μM, 6 hr), and Brefeldin A #9972 (300 ng/ml, last 5 hr of stimulation), using IL-4 (D19A10) Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of recombinant Human Interleukin-4 (hIL-4) #8919 using IL-4 (D19A10) Rabbit mAb.
Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left) or treated (right) with TPA #4174 (40 nM, 6 hr), Ionomycin #9995 (2 μM, 6 hr) and Brefeldin A #9972 (1 μg/ml, last 5 hr of stimulation), using a CD3 antibody and IL-4 (D19A10) Rabbit mAb. Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 647 Conjugate) #4414 was used as a secondary antibody. Analysis was performed on cells in the lymphocyte gate.