Preparation And Storage
Recommended Assay Procedures
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.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
- 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.
Companion Products
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.
This antibody is routinely tested by ELISA Capture. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
Development References (8)
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Abrams JS, Roncarolo MG, Yssel H, Andersson U, Gleich GJ, Silver JE. Strategies of anti-cytokine monoclonal antibody development: immunoassay of IL-10 and IL-5 in clinical samples. Immunol Rev. 1992; 127:5-24. (Clone-specific: ELISA). View Reference
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Lucin P, Pavic I, Polic B, Jonjic S, Koszinowski UH. Gamma interferon-dependent clearance of cytomegalovirus infection in salivary glands. J Virol. 1992; 66(4):1977-1984. (Clone-specific: Neutralization). View Reference
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Mo XY, Sarawar SR, Doherty PC. Induction of cytokines in mice with parainfluenza pneumonia. J Virol. 1995; 69(2):1288-1291. (Clone-specific: ELISA). View Reference
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Sadick MD, Heinzel FP, Holaday BJ, Pu RT, Dawkins RS, Locksley RM. Cure of murine leishmaniasis with anti-interleukin 4 monoclonal antibody. Evidence for a T cell-dependent, interferon gamma-independent mechanism. J Exp Med. 1990; 171(1):115-127. (Clone-specific: Neutralization). View Reference
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Sarawar SR, Sangster M, Coffman RL, Doherty PC. Administration of anti-IFN-gamma antibody to beta 2-microglobulin-deficient mice delays influenza virus clearance but does not switch the response to a T helper cell 2 phenotype. J Immunol. 1994; 153(3):1246-1253. (Clone-specific: ELISA). View Reference
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Spitalny GL, Havell EA. Monoclonal antibody to murine gamma interferon inhibits lymphokine-induced antiviral and macrophage tumoricidal activities. J Exp Med. 1984; 159(5):1560-1565. (Clone-specific: Neutralization). View Reference
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Stevenson MM, Tam MF, Belosevic M, van der Meide PH, Podoba JE. Role of endogenous gamma interferon in host response to infection with blood-stage Plasmodium chabaudi AS. Infect Immun. 1990; 58(10):3225-3232. (Clone-specific: Neutralization). View Reference
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Yang X, HayGlass KT. A simple, sensitive, dual mAb based ELISA for murine gamma interferon determination: comparison with two common bioassays. J Immunoassay. 1993; 14(3):129-148. (Clone-specific: ELISA). View Reference
Please refer to Support Documents for Quality Certificates
Global - Refer to manufacturer's instructions for use and related User Manuals and Technical data sheets before using this products as described
Comparisons, where applicable, are made against older BD Technology, manual methods or are general performance claims. Comparisons are not made against non-BD technologies, unless otherwise noted.
For Research Use Only. Not for use in diagnostic or therapeutic procedures.
