Preparation And Storage
Recommended Assay Procedures
ELISA Detection: The biotinylated JES1-5A10 antibody (Cat. No. 554491) is useful as a detection antibody for a sandwich ELISA for measuring human IL-5 protein levels. Biotinylated JES1-5A10 anti-human IL-5 antibody can be paired with the purified TRFK5 antibody (Cat. No. 554393) or the purified JES1-39D10 antibody (Cat. No. 554488) as the capture antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. Biotinylated JES1-5A10 anti-human IL-5 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 human IL-5 ranging from ~2,000 to 15 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.
Note 1: 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, IFN-γ, MCP-1, TCA-3, TNF; human IL-1α, IL-1β, IL-2, IL-3, IL-4, 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 2: This ELISA pair is recommended primarily for measuring cytokine from experimental cell culture systems. These ELISA reagents are not recommended for measuring IL-5 in serum samples. BD OptEIA™ Human IL-5 ELISA Set (Cat. No. 555202) is specially-formulated for cytokine measurement in plasma and serum.
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 JES1-5A10 monoclonal antibody specifically binds to the human Interleukin-5 (IL-5). IL-5 is a cytokine that is also known as T-cell replacing factor (TRF), Eosinophil differentiation factor (EDF), and B cell differentiation factor I. IL-5 is produced by T cells, mast cells, and eosinophils. IL-5 controls the generation, differentiation, activation and localization of eosinophils. IL-5 also activates and promotes the differentiation of basophils. IL-5 plays an important role in regulating cell-mediated immunity against parasitic infections and some tumors. High level IL-5 production is involved in some allergic diseases including asthma. IL-5 exerts its biological effects by binding to and signaling through high-affinity heterodimeric IL-5 receptors comprised of transmembrane IL-5 receptor alpha (IL-5Rα, aka, CD125) and beta common (βc, aka, CD131) chains. The βc chain serves as the common signaling subunit for the IL-5 receptor heterodimer as well as for the heterodimeric IL-3 and GM-CSF receptors. The immunogen used to generate the JES1-5A10 hybridoma was recombinant human IL-5. The JES1-5A10 antibody has been reported to cross react with IL-5 from rhesus monkey. This is a neutralizing antibody.
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 (5)
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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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Abrams JS, Silver JE, Van Dyke RE, Gleich GI. Eosinophil-active cytokines in human disease: development and use of monoclonal antibodies to IL-3, IL-5 and GMCSF. In: Gleich GJ and Kay AB, ed. Eosinophils in Allergy and Inflammation. New York: Dekker; 1994:133-157.
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Butterfield JH, Leiferman KM, Abrams J, et al. Elevated serum levels of interleukin-5 in patients with the syndrome of episodic angioedema and eosinophilia. Blood. 1992; 79(3):688-692. (Clone-specific: ELISA). View Reference
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Denburg JA, Silver JE, Abrams JS. Interleukin-5 is a human basophilopoietin: induction of histamine content and basophilic differentiation of HL-60 cells and of peripheral blood basophil-eosinophil progenitors. Blood. 1991; 77(7):1462-1468. (Biology). View Reference
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Limaye AP, Abrams JS, Silver JE, et al. Interleukin-5 and the posttreatment eosinophilia in patients with onchocerciasis. J Clin Invest. 1991; 88(4):1418-1421. (Clone-specific: ELISA). View Reference
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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.