BD PharmingenTM Purified Rat Anti-Mouse IL-10

Description

Immunocytochemistry on stimulated cells. RBC-lysed BALB/c splenocytes were enriched in CD4+ cells by panning and were cultured for 2 days with plate bound anti-mouse CD3 and soluble anti-mouse anti-CD28 in the presence of recombinant mouse IL-2 and recombinant mouse IL-4. The cells were subsequently harvested, washed and recultured with recombinant mouse IL-2 and recombinant mouse IL-4 for an additional 3 days. Finally, the cells were harvested, washed and cultured with PMA Sigma, 5 ng/ml and ionomycin Sigma, 500 ng/ml in the presence of GolgiPlug Cat. No. 555029 for 4 hr 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 a Biotin Goat anti-Rat IgG secondary antibody Cat. No. 559286 and a horseradish peroxidase-based detection system Cat. No. 550946 Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of the Purified Rat Anti-Mouse IL-10 Cat. No. 559063 antibody was blocked by the preincubation of the purified antibody with excess recombinant mouse IL-10 protein Cat. No. 550070; data not shown . Immunocytochemistry on stimulated cells. RBC-lysed BALB/c splenocytes were enriched in CD4+ cells by panning and were cultured for 2 days with plate bound anti-mouse CD3 and soluble anti-mouse anti-CD28 in the presence of recombinant mouse IL-2 and recombinant mouse IL-4. The cells were subsequently harvested, washed and recultured with recombinant mouse IL-2 and recombinant mouse IL-4 for an additional 3 days. Finally, the cells were harvested, washed and cultured with PMA Sigma, 5 ng/ml and ionomycin Sigma, 500 ng/ml in the presence of GolgiPlug Cat. No. 555029 for 4 hr 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 a Biotin Goat anti-Rat IgG secondary antibody Cat. No. 559286 and a horseradish peroxidase-based detection system Cat. No. 550946 Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of the Purified Rat Anti-Mouse IL-10 Cat. No. 559063 antibody was blocked by the preincubation of the purified antibody with excess recombinant mouse IL-10 protein Cat. No. 550070; data not shown . Show More Immunocytochemistry on stimulated cells. RBC-lysed BALB/c splenocytes were enriched in CD4+ cells by panning and were cultured for 2 days with plate bound anti-mouse CD3 and soluble anti-mouse anti-CD28 in the presence of recombinant mouse IL-2 and recombinant mouse IL-4. The cells were subsequently harvested, washed and recultured with recombinant mouse IL-2 and recombinant mouse IL-4 for an additional 3 days. Finally, the cells were harvested, washed and cultured with PMA Sigma, 5 ng/ml and ionomycin Sigma, 500 ng/ml in the presence of GolgiPlug Cat. No. 555029 for 4 hr 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 a Biotin Goat anti-Rat IgG secondary antibody Cat. No. 559286 and a horseradish peroxidase-based detection system Cat. No. 550946 Nomarski optics, original magnification 400 X . To demonstrate specificity of staining the binding of the Purified Rat Anti-Mouse IL-10 Cat. No. 559063 antibody was blocked by the preincubation of the purified antibody with excess recombinant mouse IL-10 protein Cat. No. 550070; data not shown .