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Questions? Please consult our answers to frequently asked questions at http://www.ebioscience.com/faq. DescriptionThe C8.3 antibody reacts with the p40 monomer of human IL-12 and IL-23; the antibody preferentially binds to monomeric p40. Interleukin-12 (IL-12) is a heterodimeric 70 kD (p70) cytokine composed of two covalently linked, glycosylated chains, 40kD (p40) and 35-kD (p35). IL-12 is mainly produced by monocytes, macrophages, and dendritic cells in response to bacterial products such as lipopolysaccharides (LPS), to intracellular pathogens or upon interaction with activated T cells. IL-12 was originally discovered because of its ability to induce interferon-γ production, cell proliferation, and cytotoxicity mediated by natural killer cells and T cells. It is now established that IL-12 also plays a key role in the development of Th1 responses, leading to IFN-γ and IL-2 production. These cytokines can in turn promote T-cell responses and macrophage activation. The p40 and p35 subunits by themselves have no IL-12 bioactivity, though the p40 homodimer has been shown to bind the IL-12 receptor and to be an antagonist of IL-12 p70. Free p40 is typically secreted in vast excess of IL-12 p70 by cells co-expressing both the p35 and p40 subunits. The p40 subunit of IL-12 has been shown to have extensive amino acid sequence homology to the extracellular domain of the IL-6 receptor, while the p35 subunit shows some homology to IL-6 and G-CSF. Applications ReportedFor research use only, not for diagnostic or therapeutic use. The C8.3 antibody has been reported for use in ELISA. Applications TestedThe IL-12/IL-23 p40 monomer ELISA using clones C8.3 and C8.6 preferentially measures p40 monomer, exhibiting less than 1.0% cross reactivity with IL-12 (p35p40) and IL-23 (p19p40). For this assay, the (p40 monomer-specific) purified C8.3 antibody (14-7127) is used as the capture antibody in combination with the biotinylated C8.6 antibody (13-7129) for detection and recombinant human IL-12/IL-23 p40 (39-8128) as the standard. A standard curve consisting of doubling dilutions of the recombinant standard over the range of 10,000 pg/ml - 80.0 pg/ml should be included in each ELISA plate. A suitable range of concentrations of this antibody for ELISA capture is 1-4 µg/ml. References
D’Andrea, A., et al. 1992. Production of natural killer cell stimulatory factor by peripheral blood mononuclear cells. J. Exp. Med. 176: 1387-1398. D’Andrea, A., et al. 1993. Interleukin 10 inhibits human lymphocyte interferon gamma production by suppressing natural killer cell stimulatory factor synthesis in accessory cells. J. Exp. Med. 178: 1041-1048. Chen, L., et al. 1997. Eradication of murine bladder carcinoma by intratumor injection of a bicistronic adenoviral vector carrying cDNAs for the IL-12 heterodimer and its inhibition by the IL-12 p40 subunit homodimer. J. Immunol. 159: 351-359. Ling, P., et al. 1995. Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity. J. Immunol. 154: 116-127. Russell, T., et al. 2003. IL-12 p40 homodimer-dependent macrophage chemotaxis and respiratory viral inflammation are mediated through IL-12 receptor beta1. J. Immunol. 171: 6866-6874. Related ProductsCat. 00-0000 eBioscience 10X ELISA Coating Buffer Cat. 44-2404 Nunc MaxiSorp™ flat-bottom 96 well plate Cat. 18-4100 Avidin Horse Radish Peroxidase (AV-HRP) Cat. 00-4202 eBioscience ELISA Diluent Solution, 5X Cat. 00-4203 Super AquaBlue ELISA Substrate Cat. 14-4714 Affinity Purified Mouse IgG1, K Isotype Control Cat. 14-8128 Recombinant Human IL-12 (Interleukin-12) p40 monomer Cat. 14-8129 Recombinant Human IL-12 (Interleukin-12, IL12) p70 |
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