Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The OX-81 antibody is useful as a capture antibody in a sandwich ELISA for measuring rat IL-4 protein levels in conjunction with biotinylated mouse anti-rat IL-4 monoclonal antibody (Cat. No. 555090) as detection antibody and recombinant rat IL-4 protein (Cat. No. 555107) as the standard. The OX-81 antibody should be pretitrated in the range of 1.0 to 4.0 µg/ml to determine its optimal concentration for ELISA. To obtain linear standard curves, doubling dilutions of recombinant rat IL-4 protein (e.g., ranging from ~5,120 to 5 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: This ELISA pair shows no cross-reactivity with any of the cytokines or chemokines tested (e.g., rat IL-2, IL-6, IL-10, GM-CSF, IFN-γ, 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; 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).
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 rat IL-4 in serum or plasma our rat IL-4 BD OptEIA™ Set (Cat. No. 555198) is specially formulated and recommended.
Immunofluorescent Staining and Flow Cytometric Analysis: The PE conjugated OX-81 antibody (Cat. No. 555082) is useful for immunofluorescent staining and flow cytometric analysis to identify and enumerate IL-4 producing cells.
A useful control for demonstrating specificity of staining is either of the following: 1) pre-block the fluorochrome-conjugated OX-81 antibody with ligand (e.g., recombinant rat IL-4, Cat No. 555107) prior to staining, or 2) pre-block the fixed/permeabilized cells with unlabelled OX-81 antibody (Cat. No. 555080) prior to staining. The intracellular staining technique and use of blocking controls are described in detail by C. Prussin and D. Metcalfe.
Neutralization: This antibody can be used for in vitro or in vivo neutralization of rat IL-4 bioactivity. The no azide/low endotoxin (NA/LE) format of the OX-81 clone is recommended for in vitro or in vivo assays (Cat. No. 555078).
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 OX-81 antibody reacts with rat interleukin-4 (IL-4). The immunogen used to generate this hybridoma was recombinant rat IL-4 protein. 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 (4)
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Prigent P, Saoudi A, Pannetier C, et al. Mercuric chloride, a chemical responsible for T helper cell (Th)2-mediated autoimmunity in brown Norway rats, directly triggers T cells to produce interleukin-4. J Clin Invest. 1995; 96(3):1484-1489. (Clone-specific: Cell separation, Neutralization). 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
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Ramirez F, Fowell DJ, Puklavec M, Simmonds S, Mason D. Glucocorticoids promote a TH2 cytokine response by CD4+ T cells in vitro. J Immunol. 1996; 156(7):2406-2412. (Immunogen: Neutralization). View Reference
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Saoudi A, Simmonds S, Huitinga I, Mason D. Prevention of experimental allergic encephalomyelitis in rats by targeting autoantigen to B cells: evidence that the protective mechanism depends on changes in the cytokine response and migratory properties of the autoantigen-specific T cells. J Exp Med. 1995; 182(2):335-344. (Clone-specific: Neutralization). 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.
Refer to manufacturer's instructions for use and related User Manuals and Technical Data Sheets before using this product 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.
