The NIB42 antibody reacts with human interferon-gamma (IFN-γ). The immunogen used to generate the NIB42 hybridoma was recombinant human IFN-γ. This is a neutralizing antibody.
ELISA Capture: The purified NIB42 antibody (Cat. No. 551221) is useful as a capture antibody for a sandwich ELISA for measuring human IFN-γ protein levels. The purified NIB42 antibody can be paired with the biotinylated 4S.B3 antibody (Cat. No. 554550) as the detection antibody, with recombinant human IFN-γ (Cat. No. 554617 or Cat. No. 554616) as the standard. Purified NIB42 antibody should be titrated 2.0 - 6.0 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of IFN-γ standard ranging from ~2,000 to 15 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. Note 1: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for assay of serum or plasma samples. For measuring human IFN-γ in serum or plasma our BD OptEIA™ ELISA Set (Cat. No. 555142) or BD OptEIA™ ELISA Kit (Cat. No. 550612) are specially formulated and recommended. Note 2: This ELISA pair shows no cross-reactivity with any of the cytokines or chemokines tested (eg 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, lymphotactin, MCP-1, MCP-2, MIP-1α, MIP-1β, NT-3, PDGF-AA, sCD23, SCF, TNF, LT-α, VEGF; 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; rat IL-2, IL-4, IL-6, IL-10, GM-CSF, IFN-γ, TNF). Neutralization: The no azide/low endotoxin (NA/LE™) format of the NIB42 antibody (Cat. No. 554547) is useful for neutralization of human IFN-γ bioactivity. A suitable NA/LE mouse IgG1 isotype control to match the NIB42 antibody is the 107.3 antibody, (Cat. No. 554721). WB: The NIB42 antibody is not recommended for Western blotting.
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.
