Preparation And Storage
Recommended Assay Procedures
1. ELISA Capture: The purified JES6-1A12 antibody (Cat. No. 554424) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-2 protein levels. Purified JES6-1A12 antibody can be paired with the biotinylated JES65H4 antibody (Cat. No. 554426) as the detecting antibody, with recombinant mouse IL-2 (Cat. No. 550069) as the standard. Purified JES6-1A12 antibody should be titrated 1-4 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of mouse IL-2 ranging from ~500 to 4 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
2. IP/WB: The JES6-1A12 antibody (Cat. No. 554424) has been reported to be useful for immunoprecipitation studies and for Western blotting. Please note that this application is not routinely tested at BD Biosciences.
3. IF/Flow: For immunofluorescent staining and flow cytometric analysis, the JES6-5H4 antibody has been found useful to identify and enumerate IL-2 producing cells within mixed cell populations. Directly conjugated JES6-5H4 antibodies are especially suitable for these studies. For specific methodology, please visit the protocols section or chapter on intracellular staining in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com.
4. Neutralization: The NA/LE™ S4B6 antibody (Cat. No. 554375) is useful for neutralization of mouse IL-2 bioactivity. A suitable NA/LE rat IgG2a isotype-matched control is the R35-95 antibody (Cat. No. 554687).
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 JES6-1A12 antibody reacts with mouse interleukin-2 (IL-2). The immunogen used to generate the JES6-1A12 hybridoma was recombinant mouse IL-2. 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 (5)
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Abrams J. Immunoenzymetric assay of mouse and human cytokines using NIP-labeled anti-cytokine antibodies. Curr Protoc Immunol. 2001; 1:6.20-6.21. (Clone-specific: ELISA). View Reference
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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, Immunoprecipitation). 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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Sander B, Hoiden I, Andersson U, Moller E, Abrams JS. Similar frequencies and kinetics of cytokine producing cells in murine peripheral blood and spleen. Cytokine detection by immunoassay and intracellular immunostaining. J Immunol Methods. 1993; 166(2):201-214. (Clone-specific: ELISA). 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
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.
