The R4-6A2 antibody reacts with mouse interferon-γ (IFN-γ). The immunogen used to generate the R4-6A2 hybridoma was partially purified mouse IFN-γ protein. This is a neutralizing antibody.
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. OTHER APPLICATIONS 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.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Store undiluted at 4°C.
1. Since applications vary, each investigator should titrate the reagent to obtain optimal results.
2. Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
3. Caution: Sodium azide yields highly toxic hydrazoic acid under acidic conditions. Dilute azide compounds in running water before discarding to avoid accumulation of potentially explosive deposits in plumbing.
