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 manyfold 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 p70 homodimer. Recombinant human IL-12 p70 (Cat. No. 554613) is supplied as a frozen liquid comprised of 0.22 μm sterile-filtered aqueous buffered solution, containing bovine serum albumin and glycerol, with no preservatives. Purity is ≥ 95% as determined by non-reducing SDS-PAGE, and an absorbance assay based on the Beers-Lambert Law. The endotoxin level is ≤ 0.1 ng per μg of human IL-12 p70, as measured in a chromogenic LAL assay.
Preparation And Storage
Recommended Assay Procedures
Upon initial thawing, recombinant human IL-12 p70 (Cat. No. 554613) 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 in vitro biological assay use, carrier-protein concentrations of 0.5 - 1 mg/mL are recommended. For use as an ELISA standard, carrier-protein concentrations of 5 - 10 mg/mL are recommended. Failure to add carrier protein or store at indicated temperatures may result in a loss of activity. The product should not be diluted to less than 25 μg/mL for long term storage. Carrier proteins should be pre-screened for possible effects in each investigator's experimental system. Carrier proteins may have an undesired influence on experimental results due to toxicity, high endotoxin levels or possible blocking activity.
ELISA Standard: Recombinant human IL-12 p70 (Cat. No. 554613) can be useful as a quantitative standard for measuring human IL-12 protein levels using sandwich ELISA with the purified 20C2 antibody (Cat. No.555065) as a capture antibody and biotinylated C8.6 antibody (Cat. No. 554660) as the detection antibody. To obtain linear standard curves, investigators may want to consider using doubling dilutions of recombinant human IL-12 from 2,000 - 15 pg/mL to be included in each ELISA plate. For measuring human IL-12 in serum or plasma, investigators are highly encouraged to use the BD OptEIA™ Human IL-12 ELISA Set (Cat. No. 555171 for human IL-12 p40 and Cat. No. 555183 for human IL-12 p70) or the BD OptEIA™ Human IL-12 ELISA Kit II (Cat. No. 551116 for human IL-12 p40 and Cat. No. 559258 for human IL-12 p70).
Bioassay: Investigators are advised that the Bioassay application is not routinely tested for this material and are highly encouraged to both titrate this material and include appropriate controls in relevant experiments. An activity range of 0.2 - 5.0 x 10^8 units/mg, encompassing an
ED50= 20 - 500 pg/mL, has previously been reported using PHA/IL-2 -activated human peripheral blood mononuclear cell (PBMC) for proliferation, with a unit defined as the amount of material needed to stimulate a half-maximal response at cytokine saturation.
Product Notices
- Since applications vary, each investigator should titrate the reagent to obtain optimal results.
- Source of all serum proteins is from USDA inspected abattoirs located in the United States.
- Please refer to www.bdbiosciences.com/us/s/resources for technical protocols.
Companion Products
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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Stern AS, Podlaski FJ, Hulmes JD, et al. Purification to homogeneity and partial characterization of cytotoxic lymphocyte maturation factor from human B-lymphoblastoid cells. Proc Natl Acad Sci U S A. 1990; 87(17):6808-6812. (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.
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.
