Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified JES1-39D10 antibody (Cat. No. 554488) is useful as a capture antibody for a sandwich ELISA for measuring human IL-5 protein levels. Purified JES1-39D10 antibody can be paired with the biotinylated JES1-5A10 antibody (Cat. No. 554491) as the detection antibody, with recombinant human IL-5 (Cat. No. 554606) as the standard. The purified JES1-39D10 antibody should be titrated 1.0 - 4.0 µg/ml to determine optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human IL-5 protein ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our web site, www.bdbiosciences.com, and go to the protocols section or the chapter on ELISA in the Immune Function Handbook.
Note 1: This ELISA pair shows no cross-reactivity with any of the cytokines tested (e.g., mouseIL-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, IL9, 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 assay of serum samples. For testing human IL-5 in complex biological fluids like serum or plasma, our human IL-5 OptEIA™ sandwich ELISA set (Cat. No. 555202) is recommended.
IC/Flow: Directly conjugated- JES1-39D10 is useful for intracellular cytokine flow cytometry. Purified JES1-39D10 and recombinant human IL-5 (Cat. No. 554606) are useful specificity controls for this application.
WB: The JES1-39D10 antibody has been reported to be useful for Western blotting. 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.
Companion Products
The JES1-39D10 antibody reacts with human interleukin-5 (IL-5). The immunogen used to generate the JES1-39D10 hybridoma was COS-expressed recombinant human IL-5. This is a neutralizing antibody.
This antibody is routinely tested by ELISA. Other applications were tested at BD Biosciences Pharmingen during antibody development only or reported in the literature.
Development References (7)
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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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Elson LH, Nutman TB, Metcalfe DD, Prussin C. Flow cytometric analysis for cytokine production identifies T helper 1, T helper 2, and T helper 0 cells within the human CD4+CD27- lymphocyte subpopulation. J Immunol. 1995; 154(9):4294-4301. (Clone-specific: Flow cytometry). View Reference
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Jung T, Schauer U, Rieger C, et al. Interleukin-4 and interleukin-5 are rarely co-expressed by human T cells. Eur J Immunol. 1995; 25(8):2413-2416. (Clone-specific: Flow cytometry). 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). View Reference
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Prussin C, Metcalfe DD. Detection of intracytoplasmic cytokine using flow cytometry and directly conjugated anti-cytokine antibodies. J Immunol Methods. 1995; 188(1):117-128. (Methodology: IC/FCM Block). 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.
