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Mouse il 12

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Mouse IL-12 is a lab equipment product that measures the concentration of Interleukin-12 (IL-12) in mouse samples. IL-12 is a cytokine that plays a key role in immune response and inflammation. This product provides a tool for researchers to quantify IL-12 levels in various mouse models and experimental settings.

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Lab products found in correlation

Mouse stem cell factor, by Thermo Fisher Scientific (4 mentions) Mouse thrombopoietin, by Thermo Fisher Scientific (4 mentions) Mouse tnf α, by Thermo Fisher Scientific (3 mentions) Mouse granulocyte macrophage colony stimulating factor, by Thermo Fisher Scientific (2 mentions) Atmi ku 55933, by Selleck Chemicals (2 mentions) 2 mercaptoethanol, by Thermo Fisher Scientific (2 mentions) Glutamax, by Thermo Fisher Scientific (2 mentions) Mouse il 3, by Thermo Fisher Scientific (2 mentions) Albumax 1, by Thermo Fisher Scientific (2 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions) Mouse ifn γ, by Thermo Fisher Scientific (2 mentions) Human tgf β1, by Thermo Fisher Scientific (2 mentions) Mouse il 6, by Thermo Fisher Scientific (2 mentions) Mouse il 33, by Thermo Fisher Scientific (2 mentions) Il 15, by Immunotools (2 mentions) Mouse il 18, by MBL Life Science (2 mentions) Easysep mouse cd4 positive selection kit 2, by STEMCELL (1 mentions) Soluble anti cd3, by BioXCell (1 mentions) Mouse il 4, by BioLegend (1 mentions) Anti ifn γ, by BioXCell (1 mentions)

Close spelling variants of this product

IL-12 (341 citations) Recombinant mouse IL-12 (23 citations) IL-12 p70 mouse uncoated ELISA kit (2 citations) Anti-mouse IL-12 (2 citations) IL-12 p70 Mouse ELISA Kit (2 citations) Mouse IL-12/IL-23 total p40 ELISA Ready-Set-Go (2 citations) IL-12 p40−Mouse uncoated ELISA kit (2 citations) Mouse IL-12 ELISA Total Kit (2 citations)

14 protocols using mouse il 12

1

Hematopoietic Stem Cell Culture Protocol

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Cells were clone sorted into 96-well round-bottom plates. HSCs were cultured in S-Clone (Iwai North America Inc.) supplemented with 0.75% AlbuMAX-I (Gibco), 1x penicillin/streptomycin, 50 mM 2-mercaptoethanol (Invitrogen) and the following cytokines: 20 ng/ml mouse stem cell factor, 20 ng/ml mouse thrombopoietin, 20 ng/ml mouse IL-12. Myeloid progenitors and c-kit enriched cells were cultured in Dulbecco’s modified Eagle’s medium and F-12 medium (Gibco and Invitrogen) supplemented with 10% fetal calf serum (Hyclone and Thermo Scientific), 1x penicillin/streptomycin, 2 mM GlutaMAX, 50 mM 2-mercaptoethanol (Invitrogen), and the following cytokines: 20 ng/ml mouse stem cell factor, 20 ng/ml mouse thrombopoietin, 20 ng/ml mouse IL-3, 20 ng/ml mouse granulocyte macrophage colony-stimulating factor (all purchased from PeproTech). c-kit enriched bone marrow (2×106 cells/ml) were exposed for 4 hours to 10 μM ATMi (KU55933, Selleckchem). All cells were kept in a 5% O2 incubator.
Gutierrez-Martinez P., Hogdal L., Nagai M., Kruta M., Singh R., Sarosiek K., Nussenzweig A., Beerman I., Letai A, & Rossi D.J. (2018). Diminished apoptotic priming and ATM signalling confer a survival advantage onto aged haematopoietic stem cells in response to DNA damage. Nature cell biology, 20(4), 413-421.
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2

Hematopoietic Stem Cell Culture Protocol

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Cells were clone sorted into 96-well round-bottom plates. HSCs were cultured in S-Clone (Iwai North America Inc.) supplemented with 0.75% AlbuMAX-I (Gibco), 1x penicillin/streptomycin, 50 mM 2-mercaptoethanol (Invitrogen) and the following cytokines: 20 ng/ml mouse stem cell factor, 20 ng/ml mouse thrombopoietin, 20 ng/ml mouse IL-12. Myeloid progenitors and c-kit enriched cells were cultured in Dulbecco’s modified Eagle’s medium and F-12 medium (Gibco and Invitrogen) supplemented with 10% fetal calf serum (Hyclone and Thermo Scientific), 1x penicillin/streptomycin, 2 mM GlutaMAX, 50 mM 2-mercaptoethanol (Invitrogen), and the following cytokines: 20 ng/ml mouse stem cell factor, 20 ng/ml mouse thrombopoietin, 20 ng/ml mouse IL-3, 20 ng/ml mouse granulocyte macrophage colony-stimulating factor (all purchased from PeproTech). c-kit enriched bone marrow (2×106 cells/ml) were exposed for 4 hours to 10 μM ATMi (KU55933, Selleckchem). All cells were kept in a 5% O2 incubator.
Gutierrez-Martinez P., Hogdal L., Nagai M., Kruta M., Singh R., Sarosiek K., Nussenzweig A., Beerman I., Letai A, & Rossi D.J. (2018). Diminished apoptotic priming and ATM signalling confer a survival advantage onto aged haematopoietic stem cells in response to DNA damage. Nature cell biology, 20(4), 413-421.
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3

Isolation and Polarization of Mouse CD4+ T Cells

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CD4+ T cells were isolated from the spleen and lymph nodes using the EasySep Mouse CD4 Positive Selection Kit II (Stemcell Technologies). Naive (CD25−CD44hiCD62Llo) CD4+ T cells were sorted by flow cytometry from the bead purified CD4+ T cells. The naive CD4+ T cells were resuspended in Click’s medium (Irvine Scientific) at 1 million cells per ml, and then plated on day 0 in 24 well plates coated with goat anti-hamster IgG antibody (200 ng ml−1; MP Biomedicals) with the addition of soluble anti-CD3 (1 μg ml−1; 145-2C11) and anti-CD28 (1 μg ml−1; 37.51) from Bio X Cell. Polarizing conditions for different T helper subsets are as following: TH1: human IL-2 (100 U ml−1; PeproTech), mouse IL-12 (20 ng ml−1; PeproTech) and anti-IL-4 (5 μg ml−1; Bio X Cell); TH2: human IL-2 (100 U ml−1; PeproTech), mouse IL-4 (20 ng ml−1; Biolegend), anti-IFN-γ and anti-IL-12 (5 μg ml−1; Bio X Cell); TH17: mouse IL-6 (20 ng ml−1; Biolegend), human TGF-β (2 ng ml−1; PeproTech), anti-IFN-γ and anti-IL-12 (5 μg ml−1; Bio X Cell).
Bapat S.P., Whitty C., Mowery C.T., Liang Y., Yoo A., Jiang Z., Peters M.C., Zhang L.J., Vogel I., Zhou C., Nguyen V.Q., Li Z., Chang C., Zhu W.S., Hastie A.T., He H., Ren X., Qiu W., Gayer S.G., Liu C., Choi E.J., Fassett M., Cohen J.N., Sturgill J.L., Alexander L.E., Suh J.M., Liddle C., Atkins A.R., Yu R.T., Downes M., Liu S., Nikolajczyk B.S., Lee I.K., Guttman-Yassky E., Ansel K.M., Woodruff P.G., Fahy J.V., Sheppard D., Gallo R.L., Ye C.J., Evans R.M., Zheng Y, & Marson A. (2022). Obesity alters pathology and treatment response in inflammatory disease. Nature, 604(7905), 337-342.
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4

Cytokine Injection for Tumor Therapy

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Mouse IFN-β (catalog no. 300-02BC; Peprotech Inc.), mouse TNF-α (catalog no. 315-01 A; Peprotech Inc.), and mouse IL-12 (catalog no. 210-12; Peprotech Inc.) were reconstituted in ddH2O at 1 mg/mL, respectively, and then aliquoted the reconstituted solution or stored at −80 °C to minimize freeze-thaw cycles. Based on the average concentration of the cytokines secreted from FLICs-loaded hydrogel implants every three days for 15 days, 200 ng of IFN-β, 12 ng of TNF-α, and 40 ng of IL-12 were selected to mix with 100 μL PBS containing 5% trehalose before each experiment and subcutaneously injected at the tumor resection site of each mouse, once every three days over 15 days, five times in total.
Yu Y., Wu X., Wang M., Liu W., Zhang L., Zhang Y., Hu Z., Zhou X., Jiang W., Zou Q., Cai F, & Ye H. (2022). Optogenetic-controlled immunotherapeutic designer cells for post-surgical cancer immunotherapy. Nature Communications, 13, 6357.
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5

ELISA-based Cytokine Quantification in Dendritic Cells

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Supernatants were collected from stimulated DCs (16 hrs), and cytokine production was measured by ELISA according to standard protocol. Capture and detection antibody pairs for IL-6 (MP5-20F3 and MP5-32C11), IL-10 (JES5-2A5 and SXC-1), and IL-12 (C15.6 and C17.8) were purchased from BD Biosciences. Recombinant mouse IL-6 (R&D), mouse IL-10 (R&D), and mouse IL-12 (Peprotech) were used as standards. Cytokine-antibody complexes were visualized by the addition of Tetramethyl Benzidine (TMB) solution (Life Technologies), and color development was stopped by the addition of TMB stop solution (Life Technologies). Absorbance at 492 nm was measured on a microplate reader (Biotek).
Speck S., Lim J., Shelake S., Matka M., Stoddard J., Farr A., Kuchroo V, & Laouar Y. (2014). TGF-β Signaling Initiated in Dendritic Cells Instructs Suppressive Effects on Th17 Differentiation at the Site of Neuroinflammation. PLoS ONE, 9(7), e102390.
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6

CD8+ T-cell Proliferation Assay

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Splenocytes from the mice were harvested and red blood cells were removed. The CD8+ T-cells were then sorted using a magnetic-activated cell sorting kit (CD8+ T-cell isolation kit II, Miltenyi Biotech, Bergisch Gladbach, Germany). CD8+ T-cells (1 × 106 cells/mL) were cocultured with 100 μL of PBS containing a range of cardiac myosin heavy chain-α (amino acids 334–352 peptide, Cosmo Bio Co., Ltd., Koto-Ku, Tokyo, Japan) or Influenza HA (46–54) peptide (FMYSDFHFI) (M&S TechnoSystems, Inc., Osaka-city, Osaka, Japan) at a concentration of 5 or 10 μg/mL, mouse interleukin 2 (IL-2) at a concentration of 10 or 50 IU/mL, or mouse IL-12 at a concentration of 10 or 20 ng/mL (PeproTech, Shanghai, China) for 48 h. The cultured cells were harvested and the CD8+ T-cell proliferation was assessed using the BrdU cell proliferation ELISA kit (Abcam, Cambridge, MA, USA). The supernatant was collected and analyzed for interferon-gamma (IFN-γ) release.
Hayashi T., Ichikawa M, & Konishi I. (2022). Spontaneous Myocarditis in Mice Predisposed to Autoimmune Disease: Including Vaccination-Induced Onset. Biomedicines, 10(6), 1443.
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7

Th1 Polarization of Mouse Naive CD4+ T Cells

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Naive CD4+ T cells were purified from the spleens of 7–8-week-old female mice using a CD4+CD62+ T cell Isolation Kit II (Miltenyi). Mouse naive CD4+ T cells were cultured at 106/ml in RPMI 1640 medium containing 10 mM Hepes, 10% fetal bovine serum, 2 mM L-glutamine, and antibiotics, including 50 μM 2-ME, and polarized under Th1 conditions for 3 days with plate-bound anti-CD3 (2 µg/ml), soluble anti-CD28 (1 µg/ml, PharMingen), 10 ng/ml mouse IL-12 (PeproTech), and 10 μg/ml of anti-mIL-4 antibody36 (link).
Ren M., Kazemian M., Zheng M., He J., Li P., Oh J., Liao W., Li J., Rajaseelan J., Kelsall B.L., Peltz G, & Leonard W.J. (2020). Transcription factor p73 regulates Th1 differentiation. Nature Communications, 11, 1475.
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8

T Cell Activation and Differentiation

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Spleens were harvested from C57BL/6 mice, single-cell suspensions were prepared, and T cells were activated in culture with 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28. Cells were split 1:2 with fresh media containing 7.5 ng/mL recombinant IL-2 daily starting 2 days after initial culture. After 5 days, the activated T cells were plated at 2 × 105 cells/well in 200 μL media containing mouse IL-7 at 10 ng/mL in 96-well U-bottomed plates. Where indicated, cultures were supplemented with the following cytokines (all from Peprotech): human TGF-β1 at 10 ng/mL, mouse IL-1 at 100 ng/mL, mouse IL-6 at 100 ng/mL, mouse IL-12 at 10 ng/mL, mouse IL-33 at 100 ng/mL, mouse TNF-α at 125 ng/mL, mouse IFN-γ at 10 ng/mL. Cells were then incubated for 72 hours before staining for flow cytometric analysis.
Bromley S.K., Akbaba H., Mani V., Mora-Buch R., Chasse A.Y., Sama A, & Luster A.D. (2020). CD49a Regulates Cutaneous Resident Memory CD8+ T Cell Persistence and Response. Cell reports, 32(9), 108085.
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9

Cytokine-stimulated IFN-γ Production in NK Cells

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For cytokine stimulation experiments, 2 × 10 4 mouse NK cells were stimulated for 4 h in CR-10 (RPMI 1640 + 25 mM HEPES (Gibco-15630-080) + 10% FBS, 1% l-glutamine, 1% 200 mM sodium pyruvate, 1% MEM-NEAA, 1% penicillin-streptomycin, 0.5% sodium bicarbonate and 0.01% 55 mM 2-mercaptoethanol), brefeldin A (1:1,000 dilution; BioLegend) and monensin (2 μM; BioLegend) with or without recombinant mouse IL-15 (50 ng ml -1 ; PeproTech), mouse IL-12 (20 ng ml -1 ; PeproTech) and/or recombinant mouse IL-18 (10 ng ml -1 ; BioLegend, 767002). Cells were cultured in CR-10 medium alone as a negative control (no treatment). The absolute number of IFN-γ-producing NK cells were determined by acquiring and counting individual cells with an Attune NxT after intracellular flow cytometry staining to determine cell count of IFN-γ + of the 2 × 10 4 cells plated per condition. For plate-bound antibody stimulation experiments, 2 × 10 4 isolated NK cells for each condition were stimulated with 4 mg ml -1 precoated antibody against NK1.1 (PK136) for 4 h in complete medium containing brefeldin A (1:1,000 dilution; BioLegend) and monensin (2 μM; BioLegend). Cells were cultured in medium alone as a negative control (no treatment).
Cheng M.I., Li J.H., Riggan L., Chen B., Tafti R.Y., Chin S., Ma F., Pellegrini M., Hrncir H., Arnold A.P., O'Sullivan T.E, & Su M.A. (2023). The X-linked epigenetic regulator UTX controls NK cell-intrinsic sex differences. Nature immunology, 24(5).
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10

T Cell Activation and Differentiation

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Spleens were harvested from C57BL/6 mice, single-cell suspensions were prepared, and T cells were activated in culture with 1 μg/mL anti-CD3 and 1 μg/mL anti-CD28. Cells were split 1:2 with fresh media containing 7.5 ng/mL recombinant IL-2 daily starting 2 days after initial culture. After 5 days, the activated T cells were plated at 2 × 105 cells/well in 200 μL media containing mouse IL-7 at 10 ng/mL in 96-well U-bottomed plates. Where indicated, cultures were supplemented with the following cytokines (all from Peprotech): human TGF-β1 at 10 ng/mL, mouse IL-1 at 100 ng/mL, mouse IL-6 at 100 ng/mL, mouse IL-12 at 10 ng/mL, mouse IL-33 at 100 ng/mL, mouse TNF-α at 125 ng/mL, mouse IFN-γ at 10 ng/mL. Cells were then incubated for 72 hours before staining for flow cytometric analysis.
Bromley S.K., Akbaba H., Mani V., Mora-Buch R., Chasse A.Y., Sama A, & Luster A.D. (2020). CD49a Regulates Cutaneous Resident Memory CD8+ T Cell Persistence and Response. Cell reports, 32(9), 108085.
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