The 1B1.3a antibody reacts with the extracellular region of mouse CD210 which is also known as the mouse IL-10 receptor (mIL-10R), it does not recognize the human IL-10R. The IL-10R is expressed by a variety of mouse cell types and cell lines including thymocytes, T cells, B cells, and monocytes. 1B1.3a is a neutralizing antibody and reportedly blocks the binding of human IL-10, which cross-reacts with the mouse IL-10R. The immunogen used for the generation of the 1B1.3a hybridoma was purified recombinant ligand-binding domain of mIL-10R. Mouse B220+ splenocytes were stained with purified 1B1.3a (0.25 μg, Cat. No. 559912) followed by biotinylated polyclonal goat anti-rat Ig (0.25 μg, Cat. No. 554014) and Streptavidin-PE (0.015 μg, Cat. No. 554061) in a three layer staining protocol to amplify immunofluorescent signals (left panel). In addition, cells in buffer containing sodium azide were pre-incubated on ice with recombinant mouse IL-10 (0.25 μg, Cat. No. 554585) prior to staining with 1B1.3a to determine the blocking effect of bound IL-10 protein on staining with 1B1.3a (right panel). Staining with the purified 1B1.3a antibody (filled histograms) is compared to the staining obtained with the secondary antibody alone (open histograms). Histograms in both panels are gated on the lymphocyte population as defined by light scattering characteristics and were also gated on the cell subset stained with FITC-conjugated rat-anti-mouse B220 (0.06 μg, Cat. No. 553088).
BD Pharmingen™ Purified Rat Anti-Mouse CD210 - Purified - Clone 1B1.3a - Isotype Rat IgG1, κ - Reactivity Ms - 0.5 mg
Purification
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography.
Immunogène
Purified recombinant ligand-binding domain of mIL-10R