The antibody was purified by affinity chromatography, and conjugated with biotin under optimal conditions. The solution is free of unconjugated biotin.
Storage & Handling:
The antibody solution should be stored undiluted at 4°C. Do not freeze.
Application:
ELISA Detection1,2, ELISPOT Detection
Recommended Usage:
Each lot of this antibody is quality control tested by ELISA assay. For ELISA detection applications, a concentration range of 0.5-2.0 μg/ml is recommended. 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. To obtain a linear standard curve, serial dilutions of IL-12 p70 recombinant protein ranging from 1000 to 8 pg/ml are recommended for each ELISA plate. For use as an ELISPOT detection antibody, a concentration range of 1-4 μg/ml is recommended. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
ELISA or ELISPOT Detection1,2: The biotinylated C8.6 antibody is useful as a detection antibody for an IL-12 p40 specific sandwich ELISA or ELISPOT assay, when used in conjunction with the C8.3 antibody (Cat. No. 501702/501704) as the capture antibody. The biotinylated C8.6 antibody is useful as a detection antibody for an IL-12 p70 specific sandwich ELISA or ELISPOT assay, when used in conjunction with the purified 7B12 antibody (Cat. No. 511002/511004) as the capture antibody. Biotin-F(ab') (Cat. No. 508805) is recommended for ELISA analysis of human serum and plasma samples, to prevent non-specific binding of interfering substances such as rheumatoid factor and heterophilic antibodies. Each lot of this antibody is quality control tested by ELISA assay. Neutralization1,3,4: The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of human IL-12 bioactivity (Cat. No. 508804).
Additional reported applications (for the relevant format) include: immunoprecipitation4, Western blotting1. 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. 3. Goodier, M., et al., 2000. J. Immunol. 165: 139. 4. Aragane, Y., et al., 1994. J. Immunol. 153: 5366.
Description:
The C8.6 antibody reacts with human IL-12 p40 subunit of the IL-12 p70 heterodimer and IL-23 p40 subunit of the IL-23 p19/p40, as well as p40 monomer and homodimer. The C8.6 antibody has been reported to strongly inhibit different biological activities of IL-12, (e.g., IFN-gamma induction, mitogenic effects on PHA blasts, and enhancement of NK cell-mediated cytotoxicity). The C8.6 antibody can neutralize the bioactivity of natural or recombinant IL-12.
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; p40 subunit in common with IL-23
Cellular Sources:
Dendritic cells, monocytes/macrophages, B cells, T cells
Cellular Targets:
T cells, NK cells, PBL
Receptors:
IL-12Rβ1 binds p40; dimeric with IL-12Rβ2 binds p35
Bioactivity/Activities:
IL-12 p70 (p40:p35) induces IFN-γ, TNF-a 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 human me
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.
