Description
Interleukin 12 (IL-12) is a potent regulator of cell-mediated immune responses. Biologically active IL-12 is secreted by activated B lymphocytes and macrophages as a 70 kD heterodimeric glycoprotein comprised of disulfide-bonded 35 kD (p35) and 40 kD (p40) subunits. The IL-12 p40 monomer shares amino acid sequence homology with the IL-6 receptor. It has been reported that activated PBMC produce a large excess of IL-12 p40 monomer over the bioactive p70 heterodimer. The IL-12 p40 monomer has been reported to inhibit binding of IL-12 p70 to the IL-12 receptor, but with 20X less effectiveness than the IL-12 p40 homodimer.
Formulation and Purity:
Recombinant mouse IL-12 p40 is supplied as a frozen liquid comprised of 0.22 µm sterile-filtered Dulbecco's PBS (pH 7.2) and no preservatives. The recombinant mouse IL-12, p40 is > 95% pure, as determined by SDS-PAGE, and an absorption assay based on the Beer-Lambert law. The recombinant is not tested for endotoxin or sterility.
Preparation And Storage
Store product at -80°C prior to use or for long term storage of stock solutions. Rapidly thaw and quick-spin product prior to use. Avoid multiple freeze-thaws of product. Upon initial thawing the product should be aliquoted into polypropylene microtubes and frozen at -80°C for future use. Alternatively, the product can be diluted in sterile neutral buffer containing not less than 0.5 - 10 mg/ml carrier protein** such as human or bovine albumin, aliquoted and stored at -80°C. For use as an ELISA standard we recommend carrier-protein concentrations of 5 -10 mg/ml. NOTE: Failure to add carrier protein or store at indicated temperatures may result in a loss of activity. Carrier proteins should be pre-screened for possible effects in an appropriate experimental system. Carrier proteins may affect experimental results due to toxicity, high endotoxin levels or possible blocking activity.
Recommended Assay Procedures
ELISA Standard: The recombinant mouse IL-12 p40 protein is useful as a quantitative standard for measuring mouse IL-12 protein levels in an IL-12 specific sandwich ELISA. For this assay purified anti-mouse IL-12 p40/70, clone C15.6 (Cat. No. 551219) can be used as the capture antibody when paired with biotinlyated anti-mouse IL-12 p40/p70, clone C17.8 (Cat No. 554476) for detection. To obtain standard curves, doubling dilutions of mouse IL-12 p40 ranging from ~4,000 to 30 pg/ml are recommended for inclusion in each ELISA plate. For specific methodology please visit the protocols section or chapter on ELISA in the Immune Function Handbook, both of which are located on our web site, www.bdbiosciences.com.
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.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
Development References (5)
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D'Andrea A, Aste-Amezaga M, Valiante NM, Ma X, Kubin M, Trinchieri G. Interleukin 10 (IL-10) inhibits human lymphocyte interferon gamma-production by suppressing natural killer cell stimulatory factor/IL-12 synthesis in accessory cells. J Exp Med. 1993; 178(3):1041-1048. (Biology). View Reference
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D'Andrea A, Rengaraju M, Valiante NM, et al. Production of natural killer cell stimulatory factor (interleukin 12) by peripheral blood mononuclear cells. J Exp Med. 1992; 176(5):1387-1398. (Biology). View Reference
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Ling P, Gately MK, Gubler U, et al. Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity. J Immunol. 1995; 154(1):116-127. (Biology). View Reference
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Schoenhaut DS, Chua AO, Wolitzky AG, et al. Cloning and expression of murine IL-12. J Immunol. 1992; 148(11):3433-3440. (Biology). View Reference
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Zeh HJ III, Tahara H, Lotze MT. Interleukin-12. In: Thomson AW, ed. The Cytokine Handbook. London: Academic Press Limited; 1994:239-256.
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.
