BD PharmingenTM Purified Rat Anti-Human IL-10

Description

Immunohistochemical analysis of human IL-10 expression on human peripheral mononuclear cells. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured for 2 days with plate bound anti-human CD3 and soluble anti-human CD28 in the presence of recombinant human IL-2 and recombinant human IL-4. The cells were subsequently harvested, washed and recultured with recombinant human IL-2 and recombinant human IL-4 for an additional 3 days. Finally, the cells were harvested, washed and stimulated with PMA Sigma and ionomycin Sigma in the presence of GolgiStop Cat. No. 554724 for 4 hours at 37C. The activated cells were harvested and the presence of IL-10 producing cells was detected by immunocytochemistry using a three-step staining procedure that employs Biotin Goat Anti-Rat Ig Cat. No. 559286 and Anti-Rat Ig HRP Detection Kit Cat. No. 551013 see protocol below Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of Purified Rat Anti-Human IL-10 Cat. No. 559076 antibody was blocked by the preincubation of the purified antibody with excess recombinant human IL-10 protein Cat. No. 554611; data not shown . Immunohistochemical analysis of human IL-10 expression on human peripheral mononuclear cells. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured for 2 days with plate bound anti-human CD3 and soluble anti-human CD28 in the presence of recombinant human IL-2 and recombinant human IL-4. The cells were subsequently harvested, washed and recultured with recombinant human IL-2 and recombinant human IL-4 for an additional 3 days. Finally, the cells were harvested, washed and stimulated with PMA Sigma and ionomycin Sigma in the presence of GolgiStop Cat. No. 554724 for 4 hours at 37C. The activated cells were harvested and the presence of IL-10 producing cells was detected by immunocytochemistry using a three-step staining procedure that employs Biotin Goat Anti-Rat Ig Cat. No. 559286 and Anti-Rat Ig HRP Detection Kit Cat. No. 551013 see protocol below Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of Purified Rat Anti-Human IL-10 Cat. No. 559076 antibody was blocked by the preincubation of the purified antibody with excess recombinant human IL-10 protein Cat. No. 554611; data not shown . Show More Immunohistochemical analysis of human IL-10 expression on human peripheral mononuclear cells. PBMC were isolated from human peripheral blood by density gradient centrifugation and were cultured for 2 days with plate bound anti-human CD3 and soluble anti-human CD28 in the presence of recombinant human IL-2 and recombinant human IL-4. The cells were subsequently harvested, washed and recultured with recombinant human IL-2 and recombinant human IL-4 for an additional 3 days. Finally, the cells were harvested, washed and stimulated with PMA Sigma and ionomycin Sigma in the presence of GolgiStop Cat. No. 554724 for 4 hours at 37C. The activated cells were harvested and the presence of IL-10 producing cells was detected by immunocytochemistry using a three-step staining procedure that employs Biotin Goat Anti-Rat Ig Cat. No. 559286 and Anti-Rat Ig HRP Detection Kit Cat. No. 551013 see protocol below Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of Purified Rat Anti-Human IL-10 Cat. No. 559076 antibody was blocked by the preincubation of the purified antibody with excess recombinant human IL-10 protein Cat. No. 554611; data not shown .