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Each lot of this antibody is quality control tested by ELISA assay. For ELISA capture applications, a concentration range of 1-4 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.
COA:
Application Notes:
ELISA or ELISPOT Capture1,2,4,6,8,10: The purified C15.6 antibody is useful as the capture antibody in a sandwich ELISA or ELISPOT assay, when used in conjunction with the biotinylated C17.8 (Cat. No. 505302) antibody as the detecting antibody. This assay detects p40 as monomer, homodimer, or heterodimer complexed with p35. The LEAF™ purified antibody is suggested for ELISPOT capture. Additional reported applications (for the relevant formats) include: immunohistochemical staining9 of paraformaldehyde-fixed saponin-treated frozen tissue sections, immunoprecipitation3, Western blotting3, and immunocytochemistry. Note: For testing mouse IL-12 p40 (monomer, dimer, heteromer) 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. Gately M. 1995. Curr. Prot. Immunol. John Wiley and Sons, New York. Unit 6.16. 5. Macatonia S, et al. 1995. J. Immunol. 154:5071. 6. O'Connell PJ, et al. 2006. Blood 107:1010. 7. Akilov OE, et al. 2007. J. Leukocyte Biol. 2007;10.1189/jlb.0706439. 8. Dzhagalov I, et al. 2007. J. Immunol. 178:2113. 9. Huang LY, et al. 2001. J. Immunol. 167:1423. 10. Xu G, et al. 2007. J. Immunol. 179:5358. PubMed 11. Charles N, et al. 2010. Nat. Med. 16:701. (FC) PubMed
The C15.6 antibody reacts with mouse IL-12 p40 subunit of the IL-12 p70 and IL-23 p40 subunit of the IL-23 p19/p40, as well as p40 monomer and homodimer, or heterodimer. The C15.6 antibody can not 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 ML, et al. 2002 J. Immunol. 168:5448. 9. Lankford, CS, et al. 2003 J. Leukocyte Biol. 73:49.
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