ELISA Capture: The purified BVD2-23B6 antibody (Cat. No. 554502) is useful as a capture antibody in a sandwich ELISA for measuring human GM-CSF protein levels. The purified BVD2-23B6 antibody can be paired with the biotinylated BVD2-21C11 antibody (Cat. No. 554505) as the detection antibody, with recombinant human GM-CSF (Cat. No. 550068) as the standard. Purified BVD2-23B6 antibody should be titrated 2 - 6 µ/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of GM-CSF standard ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate.
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-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, 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).
This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum samples. For testing human GM-CSF in complex biological fluids such as serum or plasma, our Human GM-CSF BD OptEIA™ ELISA Set (Cat. No. 555126) is recommended.
Neutralization: The NA/LE format of the BVD2-23B6 antibody (Cat. No. 554501) is useful for neutralization of human GM-CSF bioactivity. A suitable NA/LE. rat IgG2a isotype control is the R35-95 antibody, (Cat. No. 554687).
IP/WB: The BVD2-23B6 antibody (e.g., Cat. No. 554502) has been reported to be useful for immunoprecipitation studies. For Western blotting, a concentration of 1 µ/ml has been found to enable visualization of ≤ 100 ng/lane of recombinant human GM-CSF (e.g., Cat. No. 550068), under reducing conditions.