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Il 12

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IL-12 is a cytokine that plays a key role in the regulation of the immune system. It is a heterodimeric protein composed of two subunits, p35 and p40, and is primarily produced by antigen-presenting cells such as dendritic cells and macrophages. IL-12 is involved in the activation and differentiation of T cells and natural killer cells, and it promotes the production of interferon-gamma, a crucial cytokine in the Th1 immune response.

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Lab products found in correlation

Anti cd3, by BioXCell (5 mentions) Il 12, by BioLegend (3 mentions) Anti cd28, by BioXCell (3 mentions) Soluble anti cd28 abs, by BioXCell (2 mentions) Anti il 4 ab, by BioXCell (2 mentions) Soluble anti cd28 ab, by BioLegend (2 mentions) Anti ifn γ, by BioXCell (2 mentions) Tgf β, by Thermo Fisher Scientific (2 mentions) Il 21, by Thermo Fisher Scientific (1 mentions) Ifn γ, by BioXCell (1 mentions) Tgf β, by BioXCell (1 mentions) Interleukin 6 (il 6), by BioXCell (1 mentions) Interleukin 4 (il 4), by BioXCell (1 mentions) Interleukin 6 (il 6), by Cell Signaling Technology (1 mentions) Il 23, by Thermo Fisher Scientific (1 mentions) Easysep mouse naive cd4 t cell isolation kit, by STEMCELL (1 mentions) Ifn γ, by R&D Systems (1 mentions) Interleukin 4 (il 4), by R&D Systems (1 mentions) Il 12, by R&D Systems (1 mentions) Penicillin, by GE Healthcare (1 mentions)

Close spelling variants of this product

Anti-IL-12 Anti-mouse IL-12 (clone C17.8) Anti-mouse IL-12 p40 Anti-mouse IL-12 Anti-mouse IL-12/IL-23 p40

5 protocols using il 12

1

In Vitro Activation and Polarization of CD4+ T Cells

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For in vitro activation, naïve CD4+ T-cells were cultured in R10 medium and stimulated with plate-bound anti-CD3 (0.2 μg/ml, 0.5 μg/ml, 1.0 μg/ml) (Clone 145-2C11, BioX Cell, West Lebanon, NH) and soluble anti-CD28 Abs (1.0 μg/ml) (Clone 37.51, BioX Cell, West Lebanon, NH) for 24 and 48 h. For Th1 cell polarization, cells were stimulated with plate-bound anti-CD3 (1.0 μg/ml) and soluble anti-CD28 Abs (1.0 μg/ml) in the presence of IL-12 (10 ng/ml, Biolegend) and anti-IL-4 Ab (10 μg/ml) (Clone 11B11, BioX Cell, West Lebanon, NH) for 72 h. For phospho-STAT4 detection, cells were activated with plate-bound anti-CD3 (1.0 μg/ml) and soluble anti-CD28 (1.0 μg/ml) for 24 h, then stimulated with IL-12 (10 ng/ml) for the indicated time points.
Yang W., Gibson S.A., Yan Z., Wei H., Tao J., Sha B., Qin H, & Benveniste E.N. (2020). Protein Kinase 2 (CK2) Controls CD4+ T-cell Effector Function in the Pathogenesis of Colitis. Mucosal immunology, 13(5), 788-798.
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2

Differentiation of Naive CD4+ T Cells

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Spleen cells were excised from mice and single-cell suspensions of splenocytes were prepared in RPMI 1640 by teasing the organ through a sterile nylon mesh. Naive CD4+ T cells were purified using an EasySep mouse naive CD4+ T cell isolation kit (Stemcell Technologies, Vancouver, BC, Canada) and cultured with plate-bound anti-CD3 (5 µg/ml; Bio X Cell), anti-CD28 (5 µg/ml; Bio X Cell), and cytokines used alone, including IL-12 (R&D Systems), IFN-γ (R&D Systems), IL-1 (R&D Systems), IL-4 (R&D Systems), IL-6 (Cell Signaling Technology), IL-23 (eBioscience), and TGF-β (Invitrogen), or in combination with Abs (Bio X Cell) for Th17-promoting (2.5 ng/ml TGF-β, 20 ng/ml IL-6, 10 µg/ml anti–IFN-γ, 10 µg/ml anti–IL-4, 10 µg/ml anti–IL-2), Th1-promoting (10 ng/ml IL-12, 5 µg/ml anti–IL-4), Th2-promoting (10 ng/ml IL-4, 5 µg/ml anti–IFN-γ), or regulatory T cell (Treg)–promoting (5 ng/ml TGF-β, 5 µg/ml anti–IFN-γ, 5 µg/ml anti–IL-4) conditions. In some experiments, naive CD4+ T cells were cultured in Th17-promoting conditions and IL-21 (20 ng/ml; eBioscience) or IL-23 (50 ng/ml) was added to cultures after 24 h. Cells were harvested after 48 h for real-time PCR analysis and after 72 h for cytokine expression by flow cytometry as previously described (23 (link)).
Rus V., Nguyen V., Tatomir A., Lees J.R., Mekala A.P., Boodhoo D., Tegla C.A., Luzina I.G., Antony P.A., Cudrici C.D., Badea T.C, & Rus H.G. (2017). RGC-32 Promotes Th17 Cell Differentiation and Enhances Experimental Autoimmune Encephalomyelitis. Journal of immunology (Baltimore, Md. : 1950), 198(10), 3869-3877.
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3

Differentiation of Th1-like iTregs from Mouse CD4+ T Cells

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CD4 T cells were isolated from spleens of C57BL/6 mice using the Mojosort™ CD4 T cell isolation kit (BioLegend) and resuspended in iTreg differentiation media supplemented with 10ng/ml IL-2 (BioLegend), 10ng/ml TGFβ (BioLegend), 80 ng/ml all-trans retinoic acid (Millipore Sigma, St. Louis, MO) and 2.5 μg/mL soluble anti-CD28 (BD Biosciences). Cells were seeded into wells of a 12-well tissue culture plate pre-coated with anti-hamster IgG (Sigma-Aldrich, St. Louis, MO) plus 1 mg/ml anti-CD3 (clone 145–2C11, BioLegend) and stimulated for 7 days at 37°C. Th1-like iTregs were generated by adding 10 ng/ml IL-12 (BioLegend) on day 3 only, or on days 3 and 5 of differentiation. The cells were collected on day 7 of differentiation. Th1 cells were generated by culturing CD4 T cells with plate-bound anti-CD3ϵ (5 μg/mL) plus anti-CD28 (2.5 μg/mL) in Th1 cell differentiation media supplemented with IL-2 (10ng/ml), IL-12 (10ng/ml), and anti-mouse IL-4 (1 μg/ml; BioXcell, West Lebanon, NH). Th1 cells were collected for experiments on day 4 of differentiation. Media with a 1:1 mixture of RPMI 1640 and Dulbecco’s modified Eagle medium (GE Life Sciences, Pittsburgh, PA) supplemented with 10% FBS (Peak Serum, Wellington, CO), 2 mM L-glutamine, 1 mM sodium pyruvate, 100 U/mL penicillin, and 100 mg/mL streptomycin (GE Life Sciences) were used for cell culture.
Jadon N., Shanthalingam S., Tew G.N, & Minter L.M. (2024). PRMT5 regulates epigenetic changes in suppressive Th1-like iTregs in response to IL-12 treatment. Frontiers in Immunology, 14, 1292049.
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4

In Vitro Activation and Polarization of CD4+ T Cells

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For in vitro activation, naïve CD4+ T-cells were cultured in R10 medium and stimulated with plate-bound anti-CD3 (0.2 μg/ml, 0.5 μg/ml, 1.0 μg/ml) (Clone 145-2C11, BioX Cell, West Lebanon, NH) and soluble anti-CD28 Abs (1.0 μg/ml) (Clone 37.51, BioX Cell, West Lebanon, NH) for 24 and 48 h. For Th1 cell polarization, cells were stimulated with plate-bound anti-CD3 (1.0 μg/ml) and soluble anti-CD28 Abs (1.0 μg/ml) in the presence of IL-12 (10 ng/ml, Biolegend) and anti-IL-4 Ab (10 μg/ml) (Clone 11B11, BioX Cell, West Lebanon, NH) for 72 h. For phospho-STAT4 detection, cells were activated with plate-bound anti-CD3 (1.0 μg/ml) and soluble anti-CD28 (1.0 μg/ml) for 24 h, then stimulated with IL-12 (10 ng/ml) for the indicated time points.
Yang W., Gibson S.A., Yan Z., Wei H., Tao J., Sha B., Qin H, & Benveniste E.N. (2020). Protein Kinase 2 (CK2) Controls CD4+ T-cell Effector Function in the Pathogenesis of Colitis. Mucosal immunology, 13(5), 788-798.
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5

Differentiating Naive CD4+ T Cells into Th17 and Th1 Cells

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Spleens were collected from 6 to 8 week-old C57BL/6 female mice to obtain single-cell mixes. Naive CD4+ T cells were cultured for 72 h with 0.5 μg/mL anti-CD3 (Bioxcell) and 1 μg/mL anti-CD28 (Bioxcell). Differentiation into Th17 cells was induced by adding 10 μg/mL anti-IFN-γ (Bioxcell), 2 ng/mL TGF-β (Peprotech), 20 ng/mL IL-6 (Peprotech), and 10 ng/mL IL-1β (Peprotech). Differentiation into Th1 cells was induced by adding 10 μg/mL IL-12 (Bioxcell). CD4+ T cells were then either left unstimulated, stimulated with LPS (100 ng/mL), or with PM (400 μg/mL).
Han B., Li X., Ai R.S., Deng S.Y., Ye Z.Q., Deng X., Ma W., Xiao S., Wang J.Z., Wang L.M., Xie C., Zhang Y., Xu Y, & Zhang Y. (2022). Atmospheric particulate matter aggravates CNS demyelination through involvement of TLR-4/NF-kB signaling and microglial activation. eLife, 11, e72247.
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