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Regulatory Status Legend
Any use of products other than the permitted use without the express written authorization of Becton, Dickinson and Company is strictly prohibited.
Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified JES10-5A2 (Cat. No. 554570) is useful as a capture antibody for a sandwich ELISA for specifically measuring human IL-13 protein levels. Purified JES10-5A2 antibody can be paired with the biotinylated clone B69-2 mouse IgG1 anti-human IL-13 detection antibody (Cat. No. 555054), with recombinant human IL-13 as the standard. Purified JES10-5A2 antibody should be titrated 1.0 - 4.0 µg/ml to determine the optimal concentration for ELISA capture. To obtain linear standard curves, doubling dilutions of human IL-13 protein ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, 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., 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-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL-11, IL-12 p70, IL-12 p40, 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 or plasma samples.
Western Blot: The JES10-5A2 antibody (Cat. No. 554570) has been reported to be useful for Western blotting. Please note that this application is not routinely tested at BD Biosciences Pharmingen.
Neutralization: The NA/LE™ JES10-5A2 antibody (Cat. No. 554568) is useful for neutralization of human IL-13 bioactivity. A suitable NA/LE™ isotype control to match the JES10-5A2 antibody is the R3-34 antibody, (Cat. No. 554682).
IF/Flow: The JES10-5A2 antibody is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-13 producing cells within mixed cell populations. PE-conjugated JES10-5A2 antibody (Cat. No. 554571) is especially suitable for these studies. For specific methodology, please visit our web site, w ww.bdbiosciences.com , and go to the protocols section or the chapter on intracellular staining in the Immune Function Handbook.
Immunohistochemistry: The JES10-5A2 antibody is also reported to be useful for immunohistochemical staining studies. The purified JES10-5A2 antibody (Cat. No. 554570) may be used for the immunofluorescent staining of human IL-13, when used in conjunction with an appropriate FITC-conjugated polyclonal anti-rat Ig with no reactivity towards human Igs. For immunoenzymatic staining, the purified JES10-5A2 antibody should be detected with a biotin conjugated polyclonal anti-rat Ig, followed by an enzyme-conjugated avidin or streptavidin. The paraformaldehyde/saponin method described by J. Andersson et al. is recommended for this staining. A recommended titration range of the antibody for this staining is 2-5 µg/ml, with the optimal concentration dependent upon various factors including the type of tissue stained and the secondary reagents used. A suitable rat IgG1 isotype control for assessing the level of background staining on paraformaldehyde-fixed/saponin-permeabilized human cells or tissue is purified R3-34 antibody (Cat. No. 559072); use at comparable concentrations to antibody of interest.
Note: this antibody is not routinely tested in the IHC application.
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.
The JES10-5A2 monoclonal antibody specifically binds to human interleukin-13, IL-13. IL-13 is produced by activated T cells, mast cells and NK cells. IL-13 regulates IgE production by B cells and can suppress the cytotoxic activity of macrophages and their production of inflammatory mediators. The immunogen used to produce the JES10-5A2 hybridoma was COS-expressed recombinant human IL-13. This is a neutralizing antibody.
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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Andersson J, Abrams J, Bjork L, et al. Concomitant in vivo production of 19 different cytokines in human tonsils. Immunology. 1994; 83(1):16-24. (Clone-specific: Immunohistochemistry). View Reference
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Andersson U, Andersson J. Immunolabeling of cytokine-producing cells in tissues and in suspension. In: Fradelizie D, Emelie D, ed. Cytokine Producing Cells. Paris: Inserm; 1994:32-49.
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Litton M, Andersson J, Bjork L, Fehniger T, Ulfgren AK, Andersson U. Cytoplasmic cytokine staining in individual cells. In: Debets and Savelkoul, ed. Human Cytokine Protocols. Humana Press; 1996.
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McKenzie A, Zurawski G. Measurement of IL-13. In: Coligan, Kruisbeek, Shevak, Strober, ed. Current Protocols in Immunology. New York: John Wiley & Sons; 1994:18-19.
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.