ELISA Capture: The purified MP1-22E9 antibody (Cat. No. 554404) is useful as a capture antibody for a sandwich ELISA for measuring mouse GM-CSF protein levels. Purified MP1-22E9 antibody can be paired with the biotinylated MP1-31G6 (Cat. No. 554407) antibody as the detecting antibody, with recombinant mouse GM-CSF (Cat. No. 554586) as the standard. Purified MP1-22E9 antibody should be titrated 1-4 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse GM-CSF ranging from ~2000 to 15 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-1ß, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, IL-10, IL-12 p70, IL-15, 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, 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 GM-CSF in serum or plasma our mouse GM-CSF BD OptEIA™ set (Cat. No. 555167) is specially formulated and recommended.
Western Blot: The MP1-22E9 antibody (Cat. 554404) has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Neutralization: The NA/LE™ MP1-22E9 antibody (Cat. No. 554403) is useful for neutralization of mouse GM-CSF bioactivity.
Immunofluorescent Staining and Flow Cytometric Analysis: The MP1-22E9 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify and enumerate GM-CSF producing cells within mixed cell populations. The PE-conjugated MP1-22E9 antibody (Cat. No. 554406) is especially suitable for these experiments.