ELISA Detection: The biotinylated MQ2-39C3 antibody (Cat. No. 554546) is useful as a detection antibody for a sandwich ELISA for measuring human IL-6 protein levels. Biotinylated MQ2-39C3 antibody can be paired with the purified MQ2-13A5 (Cat. No. 554543) as the capture antibody, with recombinant human IL-6 (Cat. No. 550071) as the standard. Biotinylated MQ2-39C3 antibody should be titrated (0.5 - 2.0 µg/ml) to determine optimal concentration for ELISA detection in this system. To obtain linear standard curves, doubling dilutions of human IL-6 ranging from ~15 to 2000 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, 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-15, GM-CSF, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, IL-5, 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, LT-α, VEGF; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF).
Note 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems not serum or plasma samples. For testing human IL-6 in complex biological fluids such as serum or plasma, our human IL-6 BD OptEIA™ ELISA Set (Cat. No. 555220) and BD OptEIA™ ELISA Kit II (Cat. No. 550799) are recommended.
Western Blot: The MQ2-39C3 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.