Preparation And Storage
Recommended Assay Procedures
ELISA Detection: Biotin Rat Anti-Mouse IL-6 antibody (Cat. No. 554402) is useful as a detection antibody for a sandwich ELISA for measuring mouse IL-6 protein levels. Biotin Rat Anti-Mouse IL-6 antibody can be paired with the Purified Rat Anti-Mouse IL-6 antibody (Cat. No. 554400) as the capture antibody, with Recombinant Mouse IL-6 (Cat. No. 554582) as the standard. Biotin Rat Anti-Mouse IL-6 antibody should be pre-titered in the range of 0.5 to 2 µg/ml to determine its optimal concentration for ELISA. To obtain linear standard curves, doubling dilutions of mouse IL-6 ranging from ~2,000 to 15 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology, please visit our website, http://www.bdbiosciences.com/us/s/resources, and go to the protocols section under "ELISA and ELISPOT".
Note 1: 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. For measuring mouse IL-6 in serum or plasma our Mouse IL-6 ELISA Set (Cat. No. 555240) or Mouse IL-6 ELISA Kit (Cat. No. 550950) are specially formulated and recommended.
Note 2: 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-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-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: Biotin Rat Anti-Mouse IL-6 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.
- 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.
- For fluorochrome spectra and suitable instrument settings, please refer to our Multicolor Flow Cytometry web page at www.bdbiosciences.com/colors.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
The MP5-32C11 antibody reacts with mouse interleukin-6 (IL-6). The immunogen used to generate the MP5-32C11 hybridoma was recombinant mouse IL-6. This is a non-neutralizing antibody.
Development References (4)
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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). 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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Starnes HF Jr, Pearce MK, Tewari A, Yim JH, Zou JC, Abrams JS. Anti-IL-6 monoclonal antibodies protect against lethal Escherichia coli infection and lethal tumor necrosis factor-alpha challenge in mice. J Immunol. 1990; 145(12):4185-4191. (Biology). 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.