The antibody was purified by affinity chromatography, and conjugated with biotin under optimal conditions. The solution is free of unconjugated biotin.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C. Do not freeze.
Each lot of this antibody is quality control tested by ELISA assay. For use as an ELISA detection antibody, a concentration range of 0.25-1.0 μg/ml is recommended. For use as an ELISPOT detection antibody, a concentration range of 0.5-2.0 μg/ml is recommended. To obtain a linear standard curve, serial dilutions of IL-12 recombinant protein ranging from 250 to 2 pg/ml are recommended for each ELISA plate. It is recommended that the reagent be titrated for optimal performance for each application.
Application Notes:
ELISA Detection1,2,6-9 or ELISPOT Detection: The biotinylated C17.8 antibody is useful as a detection antibody for a sandwich ELISA or ELISPOT assay, when used in conjunction with the purified C15.6 antibody (Cat. No. 505202/505208) as capture antibody and recombinant mouse IL-12 p40 (Cat. No. 563801) or recombinant mouse IL-12 p70 (Cat. No. 563901) as the standard. Neutralization1,2,4,5,6,10: The LEAF™ purified antibody (Endotoxin <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of mouse IL-12 bioactivity in vivo and in vitro (Cat. No. 505304).
Additional reported applications (for relevant formats of this clone) include: Western blotting3, immunoprecipitation3, and intracellular immunofluorescent staining and flow cytometric analysis.
Note: For testing mouse IL-12 p40 (monomer, dimer, heterodimer) in serum or plasma, BioLegend's ELISA Max™ Sets (Cat. No. 431601 to 431606) are specially developed and recommended.
Application References:
1. Kitagaki, K., et al., 2002. Clin. Diagn. Lab Immunol. 9: 1260. 2. Reichmann, G., et al., 1999. J. Immunol. 163: 3354. 3. Wysocka, M., et al., 1995. Eur. J. Immunol. 25: 672. 4. Neucath, M., et al., 1995. J. Exp. Med. 182: 1281. 5. Cousens, LP., et al 1999 J. Exp. Med. 189:1315. 6. Gately, M. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.16. 7. O'Connell, P. J., et al., 2006. Blood 107:1010. 8. Dzhagalov, I., et al 2007. J. Immunol. 178:2113. 9. Xu, G., et al 2007. J. Immunol. 179:5358. PubMed 10. Sugimoto, K., et al 2007 Gastroenterology 133:124 PubMed
Description:
The C17.8 antibody reacts with mouse 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 C17.8 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
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.
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