ELISA Capture: The purified JES1-39D10 antibody (Cat. No. 554488) is useful as a capture antibody for a sandwich ELISA for measuring human IL-5 protein levels. Purified JES1-39D10 antibody can be paired with the biotinylated JES1-5A10 antibody (Cat. No. 554491) as the detection antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. The purified JES1-39D10 antibody should be titrated 1.0 - 4.0 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human IL-5 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, 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., mouseIL-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-6, IL-7, IL-8, IL9, 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 assay of serum samples. For testing human IL-5 in complex biological fluids like serum or plasma, our human IL-5 OptEIA™ sandwich ELISA set (Cat. No. 555202) is recommended.
IC/Flow: Directly conjugated- JES1-39D10 is useful for intracellular cytokine flow cytometry. Purified JES1-39D10 and recombinant human IL-5 (Cat. No. 554606) are useful specificity controls for this application.
WB: The JES1-39D10 antibody has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.