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Each lot of this antibody is quality control tested by intracellular immunofluorescent staining with flow cytometric analysis. Test size products are transitioning from 20 µl to 5 µl per test. Please check your vial or your CoA to find the suggested use of this reagent per million cells in 100 µl staining volume or per 100 µl of whole blood. It is recommended that the reagent be titrated for optimal performance for each application. Read more at www.biolegend.com/testsize regarding the test size change.
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.
IFN-γ-primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with with CD14 (M5E2) APC and intracellularly stained with C11.5 FITC
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.
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This data display is provided for general comparisons between formats. Your actual data may vary due to variations in samples, target cells, instruments and their settings, staining conditions, and other factors. If you need assistance with selecting the best format contact our expert technical support team.
PE anti-human IL-12/IL-23 p40 (monomer, dimer, heterodimer)
IFN-γ primed human peripheral blood monocytes were stimulated with LPS, then surface stained with CD14 APC and intracellular stained with C11.5 PE
IFN-γ-primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with with CD14 (M5E2) PE and intracellularly stained with C11.5 APC
IFN-γ-primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with with CD14 (M5E2) APC and intracellularly stained with C11.5 FITC
Alexa Fluor® 488 anti-human IL-12/IL-23 p40
IFN-g-primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with with CD14 (M5E2) PE and intracellular stained with mouse IgG1 Alexa Fluor® 488 isotype control (upper panel) or C11.5 Alexa Fluor® 488 (lower panel)
Alexa Fluor® 647 anti-human IL-12/IL-23 p40
IFN-g-primed and then LPS-stimulated (overnight) human peripheral blood monocytes surface stained with with CD14 (M5E2) FITC and intracellular stained with C11.5 Alexa Fluor® 647
Pacific Blue™ anti-human IL-12/IL-23 p40
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™
