ELISA Detection: The biotinylated BVD3-1F9 antibody (Cat. No. 554674) is useful as a detection antibody for a sandwich ELISA for measuring human IL-3 protein levels. Biotinylated BVD3-1F9 antibody can be paired with the purified BVD8-3G11 antibody (Cat. No. 554672) as the capture antibody, with recombinant human IL-3 (Cat. No. 554604) as the standard. Biotinylated BVD3-1F9 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-3 ranging from ~4,000 to 30 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.
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-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).
This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assaying serum samples. For testing human IL-3 in complex biological fluids such as serum or plasma, our BD OptEIA™ Human IL-3 ELISA Set (Cat. No. 555228) is recommended.
IC/Flow: PE- BVD3-1F9 is useful for intracellular cytokine flow cytometry (Cat. No. 554676). Purified BVD3-1F9 and recombinant human IL-3 (Cat. No. 554604) are useful specificity controls for this application.