ELISA Detection: The biotinylated JES6-5H4 antibody (Cat. No. 554426) is useful as a detection antibody for a sandwich ELISA for measuring mouse IL-2 protein levels in tissue culture supernatants. Biotinylated JES6-5H4 antibody can be paired with the purified JES6-1A12 antibody (Cat. No. 554424) as the capture antibody, with recombinant mouse IL-2 (Cat. No. 550069) as the standard. The biotinylated JES6-5H4 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 mouse IL-2 protein ranging from ~500 to 4 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our protocols or the chapter on ELISA in the Immune Function Handbook, both of which can be found at our web site, www.bdbiosciences.com.
Note: 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 mouse IL-2 in serum or plasma our BD OptEIA™ Mouse IL-2 ELISA Set (Cat. No. 555148) is specially formulated and recommended.
Note: This ELISA pair shows no cross-reactivity with any of the cytokines tested. Mouse: IL-1β, 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, TNF 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, GM-CSF, IFN-γ, TNF, LT-α, MCP-1, MCP-2, MIP-1α, MIP-1β, SCF, Lymphotactin, VEGF, G-CSF, NT-3, PDGF-AA, CD23, TCA-3 Rat: IL-2, IL-4, IL-6, IL-10, TNF-α, GM-CSF, IFN-γ.
Immunofluorescent Staining and Flow Cytometric Analysis: The JES6-5H4 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-2 producing cells within mixed cell populations. FITC, PE and APC- conjugated JES6-5H4 antibodies are available and are recommended for these studies.
IP: The purified JES6-5H4 antibody has been reported to be useful for immunoprecipitation studies. Please note that this application is not routinely tested at BD Biosciences Pharmingen.