Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified JES5-2A5 antibody (Cat. No. 551215) is useful as a capture antibody for a sandwich ELISA for measuring mouse IL-10 protein levels. Purified JES5-2A5 antibody can be paired with the biotinylated SXC-1 (Cat. No. 554423) antibody or the biotinylated JES5-16E3 (Cat No. 554465) antibody as the detecting antibody, with recombinant mouse IL-10 protein (Cat. No. 550070) as the standard. Purified JES5-2A5 antibody should be titrated (2 - 6 µg/ml) to determine the optimal concentration for ELISA capture. An appropriate range of concentrations of the mouse IL-10 recombinant protein for obtaining a linear standard curve is 2 ng/ml to 15 pg/ml mouse IL-10. For maximal sensitivity, an overnight incubation (4°C) of samples/standards with the coated capture antibody is suggested. For specific methodology, please visit the protocols section of the website or the chapter on ELISA in the Immune Function Handbook, both of which are posted on our web site, www.bdbiosciences.com. This reagent is recommended primarily for assay of cell culture supernatants. For assay of mouse IL-10 in serum or plasma, our OptEIA™ Mouse IL-10 ELISA Set (Cat. No. 555252) is recommended.
Note: This ELISA pair shows no cross-reactivity with any of the cytokines or chemokines tested (e.g., mouse IL-1β, IL-2, IL-3, IL-4, IL-5, IL-6, IL-7, IL-9, 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-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).
Western blot: The JES5-2A5 antibody (Cat. No. 551215) has been found useful for western blotting. A concentration of 1-5 µg/ml (in conjunction with the AKP-conjugated goat anti-rat Igs) has been found to enable visualization of ≤100 ng/lane of recombinant mouse IL-10 (Cat. No. 550070) under reducing conditions. Please note that this application is not routinely tested at BD Biosciences.
Neutralization: The no azide/ low endotoxin format (NA/LE) of JES5-2A5 antibody (Cat. No. 554421) is useful for neutralization of mouse IL-10 bioactivity. A suitable NA/LE rat IgG1 isotype-matched control is the R3-34 antibody (Cat. No. 554682).
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 JES5-2A5 antibody reacts with mouse IL-10. The immunogen used to generate the JES5-2A5 hybridoma was E. coli-expressed mouse IL-10. 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, 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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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
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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.
