Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 2-6 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application. To obtain a linear standard curve, serial dilutions of IL-12 (p40) recombinant protein ranging from 4000 to 30 pg/ml are recommended for each ELISA plate.
Application Notes:
ELISA or ELISPOT Capture: The purified C8.3 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated C8.6 antibody (Cat. No. 508802/508805) as the detecting antibody. The LEAF™ purified antibody is suggested for ELISPOT capture. This assay detects specifically human IL-12/IL-23 p40 protein, without interference by IL-12 p70 or IL-23 p19 proteins. Note: For testing human IL-12/IL-23 p40 (monomer, dimer) in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 430701 to 430706) are specially developed and recommended. For testing human IL-12 p70 in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 431701 to 431706) are specially developed and recommended.
Application References:
1. D'Andrea, A., et al., 1992. J. Exp. Med. 176: 1387. 2. D'Andrea, A., et al., 1993. J. Exp. Med. 178: 1041.
Description:
The C8.3 antibody reacts with human interleukin-12 and interleukin-23 (IL-12/IL-23) p40 monomer or homodimer. This antibody does not react with either the p35 subunit of IL-12 or the p19 subunit of IL-23, or the p40 protein when it is heterodimerized with p35 (the IL-12 p70) or p19 (IL-23 p60). The C8.3 antibody cannot neutralize the antagonistic activity of IL-12 p40.
Cytokine; monomer, heterodimer (p40:p35 or p40:p19) or homodimer (p40:p40)
Regulation:
Downregulated by IL-10; homodimeric p40 antagonistic to functional p70 heterodimer; p40 monomer has no function
Cellular Sources:
Dendritic cells, monocytes/macrophages, B cells, T cells
Cellular Targets:
T cells, NK cells, PBL, mature dendritic cells
Receptors:
IL-12Rβ1 binds p40 from both IL-12 and IL-23; dimeric with IL-12Rβ2 binds p35; IL-23 specific receptor subunit, IL-23R binds p19
Bioactivity/Activities:
IL-12 p70 (p40:p35) induces IFN-γ, TNF-α production in T and NK cells; costimulation of PBL proliferation; proliferation/differentiation of TH1 T lymphocytes. IL-23 (p40:p19) induces proliferation and production of IFN-γ by hu
Antigen References:
1. Fitzgerald, K., et al., Eds. 2001. The Cytokine FactsBook. Academic Press, San Diego. 2. Quesniaux, V. 1992. Research Immunol. 143:385. 3. Trinchieri, G., et al., 1992 Prog. Growth Factor Res. 4:355. 4. Trinchieri, G., et al., 1993 Immunol. Today. 14:335. 5. Oppmann, B., et al., 2000 Immunity. 13:715. 6. Aggarwal, S., et al., 2003 J. Biol. Chem.. 278:1910. 7. Parham, C., et al., 2002 J. Immunol.. 168:5699. 8. Belladonna, M.L., et al., 2002 J. Immunol.. 168:5448. 9. Lankford, C.S., et al., 2003 J. Leukoc. Biol.. 73:49.
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