ELISA Capture: Purified R4-6A2 antibody (Cat. No. 551216) is useful as a capture antibody for a mouse IFN-γ sandwich ELISA. Purified R4-6A2 antibody can be paired with biotinylated XMG1.2 antibody (Cat. No. 554410) as the detection antibody and with recombinant mouse IFN-γ protein (Cat. No. 554587) as the standard. Purified R4-6A2 antibody should be titrated 2 -6 µg/ml to determine its optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse IFN-γ ranging from ~4,000 to 30 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology please visit the protocols sections or the chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. For maximum sensitivity, an overnight incubation (4°C) of samples/standards with the coated capture antibody is suggested.
Note: 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, MCP-1, TCA-3, 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, 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: 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 IFN-γ in serum or plasma our mouse IFN-γ BD OptEIA™ Set (AN18), (Cat. No. 551866) or BD mouse IFN-γ BD OptEIA™ Kit II (Cat. No. 558258) are specially formulated and recommended.
WB: The R4-6A2 antibody has been found useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen
Neutralization: The purified R4-6A2 antibody (Cat. No. 554430) is supplied in a no azide/low endotoxin (NA/LE™) buffer. Endotoxin level as determined by LAL assay is less than 0.01 ng/µg protein. This NA/LE antibody preparation is suitable for neutralization of mouse IFN-γ bioactivity.