ELISA Detection: The biotinylated JES1-5A10 antibody (Cat. No. 554491) is useful as a detection antibody for a sandwich ELISA for measuring human IL-5 protein levels. Biotinylated JES1-5A10 anti-human IL-5 antibody can be paired with the purified TRFK5 antibody (Cat. No. 554393) or the purified JES1-39D10 antibody (Cat. No. 554488) as the capture antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. Biotinylated JES1-5A10 anti-human IL-5 antibody should be titrated 0.5 - 2.0 µg/ml to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of human IL-5 ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or 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, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, 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 measuring IL-5 in serum samples. BD OptEIA™ Human IL-5 ELISA Set (Cat. No. 555202) is specially-formulated for cytokine measurement in plasma and serum.