Mouse IL-17A Antibody

Biorbyt monoclonal anti-IL17A was used to detect IL-17A and separate Mouse CD4+ Cells by flow cytometry. Mouse CD4+ T cells were isolated from freshly dissected spleen by centrifugation in T cell separation media and selected by magnetic separation. Cells were grown on plates coated with anti-CD3 antibody, and stimulated with: 5 µg/ml anti-CD28, 10 ng/mL IL-1beta, 50 ng/mL mouse IL-6, 1 ng/mL TGFbeta1 and 10 µg/ml anti-mouse IFN gamma over 8-10 days of culture. Cells were incubated for 15-20 minutes with addition of rat anti-mouse CD4 APC at a concentration of 0.125 µg/ml, washed, fixed and permeabilized and incubated with Biorbyt Rat anti-mouse IL-17A monoclonal Antibody (orb345239) or controls as shown. Cells were washed, incubated in streptavidin conjugated PE, fixed and analyzed by Flow cytometry. Shown here are results for positive and negative controls.

Biorbyt monoclonal anti-IL-17A was used to detect IL-17A and separate Mouse CD4+ Cells by flow cytometry. Mouse CD4+ T cells were isolated from freshly dissected spleen by centrifugation in T cell separation media and selected by magnetic separation. Cells were grown on plates coated with anti-CD3 antibody, and stimulated with: 5 µg/ml anti-CD28, 10 ng/mL IL-1beta, 50 ng/mL mouse IL-6, 1 ng/mL TGFbeta1 and 10 µg/ml anti-mouse IFN gamma over 8-10 days of culture. Cells were incubated for 15-20 minutes with addition of rat anti-mouse CD4 APC at a concentration of 0.125 µg/ml, washed, fixed and permeabilized and incubated with Biorbyt Rat anti-mouse IL-17A monoclonal Antibody (orb345239) or controls as shown. Cells were washed, incubated in streptavidin conjugated PE, fixed and analyzed by Flow cytometry. Shown here are results for Biorbyt's monoclonal anti mouse IL-17A antibody (orb345239).

Western Blot showing detection of Mouse IL-17A. 100 ng of Mouse IL-17A was run on a 4-20% gel and transferred to 0.45 µm nitrocellulose. After blocking with 1% BSA-TTBS (p/n orb348540, diluted to 1X) 30 min at 20°C, Anti-Mouse IL-17A (RAT) Antibody (p/n orb345239) was used at 1:1000 in 1% BSA-TTBS over night at 4°C. Peroxidase conjugated Rabbit Anti-mouse secondary antibody (p/n orb347544) was diluted in Blocking Buffer for Fluorescent Western Blotting (p/n orb348637) at 1:40000 for 30 min at 20°C. Band indicates correct 23 kDa molecular weight position expected for Mouse IL-17A.