ELISA Detection: The biotinylated MAb11 antibody (Cat. No. 554511) can be used as a detecting antibody for a sandwich ELISA for measuring human TNF protein levels. Biotinylated MAb11 antibody can be paired with the purified MAb1 antibody (Cat. No. 551220) as the capture antibody, with recombinant human TNF (Cat. No. 554618) as the standard. This detecting antibody solution should be titrated from 0.5 - 2 µg/ml to determine its optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of TNF protein ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "ELISA and ELISPOT".
Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouse IL-1ß, 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-1ß, 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-1ß, NT3, PDGF-AA, sCD23 , SCF, 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. These ELISA reagents are not recommended for measuring serum samples. For testing human TNF in complex biological fluids like serum or plasma, our human TNF BD OptEIA™ ELISA Set (Cat. No.555212) and BD OptEIA™ ELISA Kit II (Cat. No. 550610) are recommended.
IF/Flow: The MAb11 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate TNF producing cells within mixed cell populations. FITC-conjugated MAb11 antibody (FITC-MAb11; Cat. No. 554512) and PE-conjugated MAb11 antibody (PE-MAb11; Cat. No. 554513) are recommended for these studies.