ELISA Detection: The biotinylated 4E2/MCP antibody (Cat. No. 554444) is useful as a detection antibody for a sandwich ELISA for measuring mouse MCP-1 protein levels. Biotinylated 4E2/MCP antibody can be paired with a purified antibody such as the clone 2H5 as the capture antibody, with recombinant mouse MCP-1 (Cat. No. 554590) as the standard. Biotinylated 4E2/MCP antibody should be titrated 1.0 - 4.0 µg/ml to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of mouse MCP-1 ranging from ~4,000 to 30 pg/ml are recommended for inclusion in each ELISA plate. For maximal sensitivity, an overnight incubation (4°C) of samples/standards with the coated capture antibody is suggested. For specific methodology please visit the online protocols or the chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
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, 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-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. These ELISA reagents are not recommended for assaying serum or plasma samples. For measuring mouse MCP-1 in serum or plasma the BD OptEIA™ mouse MCP-1 ELISA Set (Cat. No. 555260) is specially formulated and recommended.
Western Blot: The 4E2/MCP antibody has been found useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.