Preparation And Storage
Recommended Assay Procedures
ELISA Detection: The biotinylated JES6-5H4 antibody (Cat. No. 554426) is useful as a detection antibody for a sandwich ELISA for measuring mouse IL-2 protein levels in tissue culture supernatants. Biotinylated JES6-5H4 antibody can be paired with the purified JES6-1A12 antibody (Cat. No. 554424) as the capture antibody, with recombinant mouse IL-2 (Cat. No. 550069) as the standard. The biotinylated JES6-5H4 antibody should be titrated 0.5 - 2.0 µg/ml to determine optimal concentration for ELISA detection. To obtain linear standard curves, doubling dilutions of mouse IL-2 protein ranging from ~500 to 4 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our protocols or the chapter on ELISA in the Immune Function Handbook, both of which can be found at our web site, www.bdbiosciences.com.
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 IL-2 in serum or plasma our BD OptEIA™ Mouse IL-2 ELISA Set (Cat. No. 555148) is specially formulated and recommended.
Note: This ELISA pair shows no cross-reactivity with any of the cytokines tested. Mouse: IL-1β, 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, 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, GM-CSF, IFN-γ, TNF, LT-α, MCP-1, MCP-2, MIP-1α, MIP-1β, SCF, Lymphotactin, VEGF, G-CSF, NT-3, PDGF-AA, CD23, TCA-3 Rat: IL-2, IL-4, IL-6, IL-10, TNF-α, GM-CSF, IFN-γ.
Immunofluorescent Staining and Flow Cytometric Analysis: The JES6-5H4 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-2 producing cells within mixed cell populations. FITC, PE and APC- conjugated JES6-5H4 antibodies are available and are recommended for these studies.
IP: The purified JES6-5H4 antibody has been reported to be useful for immunoprecipitation studies. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
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.
- Use of these products to measure activation antigens expressed on mononuclear cell subsets for the purpose of monitoring immunoregulatory status can fall under one or more claims of the following patents: US Patent Nos. 5,445,939, 5,656,446, 5,843,689; European Patent No. 319,543; Canadian Patent No. 1,296,622; Australian Patent No. 615,880; and Japanese Patent No. 2,769,156.
Companion Products
The JES6-5H4 monoclonal antibody specifically binds to mouse interleukin-2 (IL-2), a multifunctional cytokine that plays pivotal roles in immunity and tolerance. It is produced by activated T cells and affects the activation, growth, proliferation and/or differentiation of various cell types including T and B lymphocytes and their precursors, LAK cells, NK cells, and monocytes/macrophages. IL-2 mediates its biological activities by binding to IL-2 receptor complexes. The intermediate affinity IL-2R is comprised of IL-2Rβ (CD122) and common gamma chain (γc; CD132) subunits, whereas the high-affinity IL-2R is comprised of IL-2Rα (CD25), IL-2Rβ, and γc subunits. The JES6-5H4 monoclonal antibody binds to IL-2 and neutralizes its biological activity.
This antibody is routinely tested by ELISA Detection. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
Development References (3)
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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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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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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.
