Western blot analysis of extracts from purified CD4 + human peripheral blood mononuclear cells, activated with anti-CD3 and anti-CD28 in regulatory T cell inducing conditions for 12 d and then 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-10 (D13A11) XP ® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower).
Western blot analysis of extracts from purified CD14 + human peripheral blood mononuclear cells, untreated (-) or treated with LPS (100 ng/mL, 16 hr; +), using IL-10 (D13A11) XP ® Rabbit mAb (upper) or β-Actin (D6A8) Rabbit mAb #8457 (lower). Brefeldin A #9972 (300 ng/mL) was added to cells after 1 hr of LPS stimulation.
Western blot analysis of recombinant Human Interleukin-10 (hIL-10) #8903 using IL-10 (D13A11) XP ® Rabbit mAb.
Flow cytometric analysis of purified CD14+ human peripheral blood mononuclear cells untreated (left) or treated with LPS (100ng/mL, 16 hr; right) using a CD14 antibody and IL-10 (D13A11) XP ® Rabbit mAb. Brefeldin A #9972 (300ng/mL) was added to untreated and treated cells after 1 hr of LPS stimulation. Anti-rabbit IgG (H+L), F(ab') 2 Fragment (Alexa Fluor ® 647 Conjugate) #4414 was used as a secondary antibody.
Flow cytometric analysis of human peripheral blood mononuclear cells, untreated (left) or treated (right) with TPA #4174 (40 nM, 5 hr), Ionomycin #9995 (2 μM, 5 hr) and Brefeldin A #9972 (1 μg/mL, last 4 hr of stimulation), using a CD3 antibody and IL-10 (D13A11) XP ® 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.