Preparation And Storage
Recommended Assay Procedures
ELISA Capture: The purified 8D4-8 antibody (Cat. No. 554515) is useful as a capture antibody for a sandwich ELISA for measuring human IL-4 protein levels. Purified 8D4-8 antibody can be paired with the biotinylated MP4-25D2 antibody (Cat. No. 554483) as the detecting antibody, with recombinant human IL-4 protein (Cat. No. 554605) as the standard. 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 Applications Manual.
Note: 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-5, IL-6, IL-7, IL-8, IL-9, IL-10, IL11, 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: 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 human IL-4 in serum or plasma our Human IL-4 BD OptEIA™ set (Cat. No. 555194) or BD OptEIA™ kit (Cat. No. 550614) are specially formulated and recommended.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- 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.
- Sodium azide is a reversible inhibitor of oxidative metabolism; therefore, antibody preparations containing this preservative agent must not be used in cell cultures nor injected into animals. Sodium azide may be removed by washing stained cells or plate-bound antibody or dialyzing soluble antibody in sodium azide-free buffer. Since endotoxin may also affect the results of functional studies, we recommend the NA/LE (No Azide/Low Endotoxin) antibody format, if available, for in vitro and in vivo use.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
The 8D4-8 monoclonal antibody reacts with human interleukin-4 (IL-4). The immunogen used to raise the 8D4-8 hybridoma was recombinant human IL-4. The 8D4-8 antibody binds to an epitope that is different than the epitope recognized by the MP4-25D2 antibody (Cat. No. 554485).
Clone 8D4-8 displays an increased amount of non-specific binding to dead cells when compared to the clone MP4-25D2. It is recommended to use a fixable viability dye in conjunction with this clone.
Development References (2)
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Bird C, Wadhwa M, Thorpe R. Development of immunoassays for human interleukin 3 and interleukin 4, some of which discriminate between different recombinant DNA-derived molecules. Cytokine. 1991; 3(6):562-567. (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). 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.