The C8.6 antibody reacts with human IL-12 p40 monomers, homodimers, and p70 heterodimers, but not p35 subunits. The immunogen used to generate the C8.6 hybridoma was the purified p40 subunit of human IL-12. The C8.6 antibody is a neutralizing antibody and has been reported to strongly inhibit three different biological activities of IL-12 (i.e., IFN-γ induction), mitogenic effects on PHA blasts, and enhancement of NK cell-mediated cytotoxicity. C8.6 has been shown to immunoprecipitate IL-23.
ELISA Detection (IL-12 p70 protein ELISA): The biotinylated C8.6 antibody (Cat. No. 554660) is useful as a detection antibody for a sandwich ELISA for specifically measuring human IL-12 p70 protein levels. Biotinylated C8.6 antibody can be paired with the purified rat anti-human IL-12 p70 specific 20C2 (Cat. No. 555065) as the capture antibody and recombinant human IL-12 p70 protein (Cat. No. 554613) as the standard. Biotinylated C8.6 antibody should be titrated (0.5 - 2.0 µg/ml) to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of human IL-12 p70 ranging from ~2000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For testing IL-12 p70 in serum or plasma BD OptEIA™ Set (Cat. No. 555183) or BD OptEIA™ Kit (Cat. No. 559258) is recommended. Note 1: This ELISA for measuring human IL-12 p70 protein levels is not interfered with by a 50-fold excess (by mass) of the human p40 monomer. This assay is specific for measuring human IL-12 p70 protein. It has been reported that activated PBMC produce an excess of free p40 protein over the p70 heterodimer. ELISA Detection (IL-12 p40 protein ELISA): The biotinylated C8.6 antibody (Cat. No. 554660) is useful as a detection antibody for specifically measuring human IL-12 p40 protein levels, with no reactivity to human IL-12 p70 protein. Biotinylated C8.6 antibody can be paired with the purified (exclusively p40 specific) C8.3 antibody (Cat. No. 551227) as the capture antibody with recombinant human IL-12 p40 (Cat. No. 554633) as the standard. For testing IL-12 (p40) in complex biological fluids like serum or plasma BD OptEIA™ Set (Cat. No. 555171) or BD OptEIA™ Kit (Cat. No. 551116) is recommended. Note 2: These ELISA pairs show no cross-reactivity with any of the cytokines or chemokines tested (e.g., human IL-1α, IL-1β, IL-2, IL-3, IL-4. IL-5, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-13, IL-15, G-CSF, GM-CSF, IFN-γ, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23, SCF, TNF, Lt-α, VEGF; mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF). Neutralization: The C8.6 antibody has been reported to be useful for neutralization of human IL-12 p70 bioactivity. A formulation that does not contain sodium azide formulation (NA/LE™) is available (Cat. No. 554659) and is recommended for functional assay. This application is not routinely tested at BD Biosciences Pharmingen.
The antibody was conjugated with biotin under optimum conditions, and unreacted biotin was removed. The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
