The antibody was purified by affinity chromatography, and conjugated with Pacific Blue™ under optimal conditions. The solution is free of unconjugated Pacific Blue™.
Concentration:
0.5 mg/ml
Storage & Handling:
The antibody solution should be stored undiluted at 4°C and protected from prolonged exposure to light. Do not freeze.
Application:
ICFC3,5
Recommended Usage:
Each lot of this antibody is quality control tested by 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. It is recommended that the reagent be titrated for optimal performance for each application.
* Pacific Blue™ has a maximum emission of 455 nm when it is excited at 405 nm. Prior to using Pacific Blue™ conjugate for flow cytometric analysis, please verify your flow cytometer's capability of exciting and detecting the fluorochrome. ** Pacific Blue™ is a registered trademark of Molecular Probes, Inc. Pacific Blue™ dye antibody conjugates are sold under license from Molecular Probes, Inc. for research use only, except for use in combination with microarrays and high content screening, and are covered by pending and issued patents.
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 <0.1 EU/μg, Azide-Free, 0.2 μm sterile-filtered) is recommended for neutralization of human IL-12 bioactivity (Cat. No. 501813).
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, C.A., et al., 2004. J. Virol. 78: 13173.
INF-γ primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with CD14 PE and intracellularly stained with C11.5 Pacific Blue™
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, M.L., et al., 2002 J. Immunol.. 168:5448. 9. Lankford, C.S., et al., 2003 J. Leukoc. Biol.. 73:49.
