ELISA Capture: IL-12 p70 ELISA: The purified 9A5 antibody (Cat. No. 554658) is useful as a capture antibody for a sandwich ELISA for specifically measuring mouse IL-12 p70 protein levels. Purified 9A5 antibody can be paired with the biotinylated C17.8 antibody (Cat. No. 554476) as the detection antibody, with recombinant mouse IL-12 p70 (Cat. No. 554592) as the standard. The purified 9A5 antibody should be titrated to determine its optimal concentration for ELISA capture (2 to 4 µg/ml). To obtain linear standard curves, doubling dilutions of mouse IL-12 ranging from ~2000 - 15 pg/ml are recommended for inclusion in each ELISA pair. For specific methodology, see BD Pharmingen's protocols or the chapter on ELISA in the Immune Function Handbook posted on the website at www.bdbiosciences.com. Caution: Use TMB substrate for higher substrate signal and higher sensitivity. For testing IL-12 p70 in serum or plasma, the OptEIA™ set (Cat. No. 555256) is recommended.
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-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF-α; 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-12 p40, 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-α, TNF-β, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF-α).