ELISA Capture: The purified C8.3 antibody (Cat. No. 551227) is useful as a capture antibody for a sandwich ELISA for specifically measuring human IL-12 p40 protein levels, without interference by IL-12 p70 protein. Purified C8.3 antibody can be paired with the biotinylated C8.6 (Cat. No. 554660) antibody as the detecting antibody, with recombinant human IL-12 p40 monomer (Cat. No. 554633) as the standard. Purified C8.3 antibody should be titrated 4 - 8 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human C8.3 ranging from ~10,000 to 80 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-12 p40, IL-15, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, 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; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF).
Note 2: This assay is specific for measuring human IL-12 p40 protein, with minimal interference by IL-12 p70 protein. IL-12 p70 protein at 10 ng/ml run in this IL-12 p40 ELISA was found to yield < 0.1% the signal produced by 10 ng/ml IL-12 p40 protein.
Note 3: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum or plasma samples. For measuring human IL-12 (p40) in serum or plasma our human IL-12 (p40) BD OptEIA™ Set (Cat. No. 555171) or BD OptEIA Kit (Cat. No. 551116) are specially formulated and recommended.