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Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. For immunofluorescent staining, the suggested use of this reagent is ≤ 0.25 µg per 106 cells in 100 µl volume. The purified C11.5 has been tested by blocking fluorochrome conjugated C11.5 for intracellular cytokine staining. In order to obtain complete blocking results, a saturated amount of purified antibody (≤ 5.0 ug/million cells) should be used for incubation with target cells, prior to staining with fluorochrome conjugated antibody. It is recommended that the reagent be titrated for optimal performance for each application.
COA:
Application Notes:
Flow Cytometry3,5: The fluorochrome-labeled C11.5 antibody is useful for intracellular immunofluorescent staining and flow cytometric analysis to identify IL-12 p40 (monomeric, heterodimeric, homodimeric) -producing cells within mixed cell populations. View intracellular cytokine staining protocol Neutralization6: The LEAF™ purified antibody (Endotoxin Additional reported applications (for the relevant formats) include: Western blotting,ELISA Capture1,2, ICFC Blocking, immunohistochemical staining4 of paraformaldehyde-fixed, saponin-treated tissue frozen sections, and immunocytochemistry. 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. Prussin C, et al. 1995. J. Immunol. Meth. 188:117. 4. Leifeld L, et al. 2002. Hepatology 36:1001. 5. Pekkari K, et al. 2001. Blood 97:3184. 6. Bolovan-Fritts CA, et al. 2004. J. Virol. 78:13173.
Description:
The C11.5 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 C11.5 antibody can neutralize the bioactivity of natural or recombinant IL-12 p70.
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-12Rß1 binds p40; dimeric with IL-12Rß2 binds p35
Bioactivity/Activities:
IL-12 p70 (p40:p35) induces IFN-?, 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-? by human memory T cells, but not n
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. Leukoc. Biol. 73:49.
*These products may be covered by one or more Limited Use Label Licenses (see the BioLegend Catalog or our website, www.biolegend.com/terms). BioLegend products may not be transferred to third parties, resold, modified for resale, or used to manufacture commercial products or to provide a service to third parties without written approval of BioLegend. By use of these products you accept the terms and conditions of all applicable Limited Use Label Licenses. Unless otherwise indicated, these products are for research use only and are not intended for human or animal diagnostic, therapeutic or commercial use.
