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Recombinant mouse il 23

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Recombinant mouse IL-23 is a cytokine that belongs to the IL-12 family. It is a heterodimeric protein composed of the p19 and p40 subunits. IL-23 plays a role in the differentiation and expansion of Th17 cells.

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Lab products found in correlation

Micrometer, by Mitutoyo (3 mentions) Nystatin, by Merck Group (1 mentions) Nocodazole, by Merck Group (1 mentions) Female c57bl 6 b6 mice, by Japan SLC (1 mentions) Pitstop 2, by Abcam (1 mentions) Anti ha antibody 12ca5, by Roche (1 mentions) Anti flag m2 antibody, by Merck Group (1 mentions) Ifn γ xmg1.2, by BioLegend (1 mentions) Recombinant mouse il 1β, by Thermo Fisher Scientific (1 mentions) Anti cd28 clone 37, by BioXCell (1 mentions) Recombinant mouse il 6, by Thermo Fisher Scientific (1 mentions) 6 formylindolo 3 2 b carbazole ficz, by Enzo Life Sciences (1 mentions) Recombinant mouse il 12, by Thermo Fisher Scientific (1 mentions) Anti il 4 mab clone 11b11, by BioXCell (1 mentions) Anti ifn γ mab clone xmg1, by BioXCell (1 mentions) Recombinant human tgf β, by R&D Systems (1 mentions) Carboxyfluorescein succinimidyl ester (cfse), by Thermo Fisher Scientific (1 mentions) Fetal bovine serum (fbs), by Gemini Bio (1 mentions)

Spelling variants of this product

IL-23 (185 citations) Recombinant IL-23 (7 citations) Recombinant mouse or human IL-23 (2 citations)

9 protocols using recombinant mouse il 23

1

Interleukin-23 Induced Mouse Skin Inflammation

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Example 28

IL-23 Injection Model of Mouse Skin Inflammation

Ears from BALB/c mice were each injected intra-dermally every other day with 150 ng of mouse recombinant IL-23 (eBiosciences) or PBS in a total volume of 25 μl. Ear swelling was measured in triplicate using a micrometer (Mitutoyo) right before each IL-23 challenge. On Day 14, mice were euthanized and ears were collected for measurement of cytokine levels, gene expression levels and hystopathological evaluation. Mice were administered 3-100 mg/kg of an RORC2 modulator or vehicle once daily orally for the duration of the study. Alternatively, the RORC2 modulator was applied topically once or twice daily using a standard formulation (EtOH:propylene glycol:dimethyl isosorbide:DMSO, 38:30:15:15) at a concentration of 0.1% to 5.0%.

References describing aspects of this assay include: Muramoto, K. et al. J. Pharmacol. Exp. Ther. 2010, 335(1), 23-31; Fridman, J. S. et al. J. Invest. Dermatol. 2011, 131(9), 1838-1844.

US10336748B2. Methyoxy-substituted pyrrolopyridine modulators of RORC2 and methods of use thereof (2019-07-02). Pfizer Inc. [US]. Inventors: Mark Edward Schnute [US], Andrew Christopher Flick [US], Peter Jones [US], Neelu Kaila [US], Scot Richard Mente [US], John David Trzupek [US], Michael L. Vazquez [US], Goran Mattias Wennerstal [SE], Li Xing [US], Edouard Zamaratski [SE], Liying Zhang [US], Rayomand J. Unwalla [US].
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2

IL-23 Induced Skin Inflammation

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Example 41

IL-23 Injection Model of Mouse Skin Inflammation

Ears from BALB/c mice were each injected intra-dermally every other day with 150 ng of mouse recombinant IL-23 (eBiosciences) or PBS in a total volume of 25 μl. Ear swelling was measured in triplicate using a micrometer (Mitutoyo) right before each IL-23 challenge. On Day 14, mice were euthanized and ears were collected for measurement of cytokine levels, gene expression levels and hystopathological evaluation. Mice were administered 3-100 mg/kg of an RORC2 modulator or vehicle once daily orally for the duration of the study. Alternatively, the RORC2 modulator was applied topically once or twice daily using a standard formulation (EtOH:propylene glycol:dimethyl isosorbide:DMSO, 38:30:15:15) at a concentration of 0.1% to 5.0%.

References describing aspects of this assay include: Muramoto, K. et al. J. Pharmacol. Exp. Ther. 2010, 335(1), 23-31; Fridman, J. S. et al. J. Invest. Dermatol. 2011, 131(9), 1838-1844.

US10385036B2. Sulfonamide-substituted indole modulators of RORC2 and methods of use thereof (2019-08-20). Pfizer Inc. [US]. Inventors: Mark Edward Schnute [US], Andrew Christopher Flick [US], Peter Jones [US], Neelu Kaila [US], Scot Richard Mente [US], John David Trzupek [US], Michael L. Vazquez [US], Goran Mattias Wennerstal [SE], Li Xing [US], Edouard Zamaratski [SE], Liying Zhang [US], Rayomand J. Unwalla [US].
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3

IL-23-Induced Mouse Skin Inflammation Model

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Example 106

IL-23 Injection Model of Mouse Skin Inflammation

Ears from BALB/c mice were each injected intra-dermally every other day with 150 ng of mouse recombinant IL-23 (eBiosciences) or PBS in a total volume of 25 μl. Ear swelling was measured in triplicate using a micrometer (Mitutoyo) right before each IL-23 challenge. On Day 14, mice were euthanized and ears were collected for measurement of cytokine levels, gene expression levels and hystopathological evaluation. Mice were administered 3-100 mg/kg of an RORC2 modulator or vehicle once daily orally for the duration of the study. Alternatively, the RORC2 modulator was applied topically once or twice daily using a standard formulation (EtOH:propylene glycol:dimethyl isosorbide: DMSO, 38:30:15:15) at a concentration of 0.1% to 5.0%.

References describing aspects of this assay include: Muramoto, K. et al. J. Pharmacol. Exp. Ther. 2010, 335(1), 23-31; Fridman, J. S. et al. J. Invest. Dermatol. 2011, 131(9), 1838-1844.

US10426135B2. Methyl- and trifluromethyl-substituted pyrrolopyridine modulators of RORC2 and methods of use thereof (2019-10-01). Pfizer Inc. [US]. Inventors: Mark Edward Schnute [US], Andrew Christopher Flick [US], Peter Jones [US], Neelu Kaila [US], Scot Richard Mente [US], John David Trzupek [US], Michael L. Vazquez [US], Goran Mattias Wennerstal [SE], Li Xing [US], Edouard Zamaratski [SE], Liying Zhang [US], Rayomand Jal Unwalla [US].
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4

Lentiviral Modulation of STAT3 and RORγt

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Following lentiviral infection, cells were seeded at a concentration of 5×106 cells/well into a 96-well culture plate and incubated with recombinant mouse IL-23 (10 ng/ml, eBioscience, Inc.) for 24 h. The protein levels of p-STAT3 and RORγt were analyzed by western blotting following the method described above. The mRNA levels of STAT3 and RORγt in lung CD4+ T cells were analyzed by RT-qPCR. The sequences of primers were as follows: Mouse STAT3 sense, 5′-TGTCTCCACTTGTCTACCT-3′ and anti-sense, 5′-CAGCACCTTCACCGTTAT-3′; mouse RORγt sense, 5′-TCTCTGCAAGACTCATCGACAAG-3′ and anti-sense, 5′-GCACAGGCTCCGGAGTTTT-3′. β-actin was used as an internal positive control for normalization of data. The results were compared between SOCS3 overexpressing group and control groups.
Ding F.M., Liao R.M., Chen Y.Q., Xie G.G., Zhang P.Y., Shao P, & Zhang M. (2017). Upregulation of SOCS3 in lung CD4+ T cells in a mouse model of chronic PA lung infection and suppression of Th17-mediated neutrophil recruitment in exogenous SOCS3 transfer in vitro. Molecular Medicine Reports, 16(1), 778-786.
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5

Intralesional IL-23 Injection in Mice

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Briefly, slc15a4feeble and C57BL/6 J mice were used at 6–8 wks of age and gender did not affect different outcomes. Mice received intralesional injections of 20 μl PBS, either alone or containing 500 ng recombinant mouse IL-23 (eBioscience), using a 26-gauge needle every other day for 16 days as previously described41 (link). Ear thickness was measured via micrometer before injection on day 0 and thereafter on days without injections.
Griffith A.D., Zaidi A.K., Pietro A., Hadiono M., Yang J.S., Davis R, & Popkin D.L. (2018). A requirement for slc15a4 in imiquimod-induced systemic inflammation and psoriasiform inflammation in mice. Scientific Reports, 8, 14451.
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6

Murine Inflammation and Immunomodulation

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Female C57BL/6 (B6) mice were purchased from Japan SLC (Hamamatsu, Japan). All experiments were conducted on 8–12-week-old mice. Recombinant mouse IL-23 was purchased from eBioscience (San Diego, CA). MaR1 was purchased from Cayman Chemical (Ann Arbor, MI). Nocodazole, nystatin, and chlorprozine were purchased from Sigma-Aldrich (Poole, UK). Pit stop2 was purchased from Abcam (Cambridge, MA). IMQ was kindly provided by Mochida Pharmaceutical Co., Ltd.
Saito-Sasaki N., Sawada Y., Mashima E., Yamaguchi T., Ohmori S., Yoshioka H., Haruyama S., Okada E, & Nakamura M. (2018). Maresin-1 suppresses imiquimod-induced skin inflammation by regulating IL-23 receptor expression. Scientific Reports, 8, 5522.
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7

Antibody Characterization for Cell Signaling

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Anti—c-Maf (M-153), anti-Hsp90 (sc-7947) and anti-Oct1 (sc-232) antibodies were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). HRP-conjugated 4G10 (anti-phosphotyrosine), HRP-conjugated goat anti-mouse and goat anti-rabbit L chain antibodies were purchased from Millipore (Billerica, MA). Anti-p-Tyr (P-Tyr-1000) antibody was purchased from Cell Signaling Technology (Danvers, MA). Anti-HA (12CA5) antibody and Protein A and G agarose beads were purchased from Roche (Mannheim, Germany). Anti-FLAG (M2) antibody was purchased from Sigma-Aldrich (St. Louis, MO). Anti-EGFP (ab290) antibody was purchased from Abcam (Cambridge, MA). Anti-mouse Ptpn22 (anti-PEP) antibody was obtained from Genentech (San Francisco, CA). Anti-Tec antibody was purchased from Upstate biotechnology. Anti-α-Tubulin 4a antibody was purchased from GeneTex. Antibodies to CD3-ε (17A2), CD28 (37.51), IL-4 (11B11), and IFNγ (XMG1.2) were purchased from BioLegend (San Diego, CA). Recombinant human IL-2 and TGF-β and recombinant mouse IL-4 and IL-6 were purchased from PeproTech (Rocky Hill, NJ). Recombinant mouse IL-23 was purchased from eBioscience (San Diego, CA).
Liu C.C., Lai C.Y., Yen W.F., Lin Y.H., Chang H.H., Tai T.S., Lu Y.J., Tsao H.W., Ho I.C, & Miaw S.C. (2015). Reciprocal Regulation of C-Maf Tyrosine Phosphorylation by Tec and Ptpn22. PLoS ONE, 10(5), e0127617.
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8

In Vitro Differentiation of Naïve T Cells

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FACS-sorted or magnetic bead-selected naïve CD4 T cells were cultured in Iscove’s Modified Dulbecco’s medium (IMDM) (Mediatech; Manassas, VA) with 10% FBS (Gemini Bio Products; West Sacramento, CA), 100 U/ml penicillin, 100 μg/ml streptomycin and 2 mM glutamine. Cells were activated with 10 μg/ml plate-bound anti-CD3 (clone X145-2C11) and anti-CD28 (clone 37.51) (Bio X Cell; West Lebanon, NH). Where indicated cultures included: 50 ng/ml recombinant mouse IL-23, 20 ng/ml recombinant mouse IL-6, 20 ng/ml recombinant mouse IL-1β, 3.5 ng/ml recombinant mouse IL-12 (all from eBioscience), 2 ng/ml recombinant human TGF-β (R&D Systems; Minneapolis, MN), 200 nM 6-formylindolo(3,2-b)carbazole (FICZ) (Enzo Life Sciences; Farmingdale, NY), 10 μg/ml anti-IFNγ mAb (clone XMG1.2), 10 μg/ml anti-IL-4 mAb (clone 11B11) (Bio X Cell). Our Th cultures conditions were: Th0: anti-IL-4 mAb, anti-IFNγ mAb; Th1: anti-IL-4 Ab, IL-12; Th17 inflammatory: anti-IL-4 mAb, anti-IFNγ mAb, IL-6, TGF-β, IL-1β, IL-23; Th17 classic: anti-IL-4 mAb, anti-IFNγ mAb, IL-6, TGF-β; and Th22: anti-IL-4 mAb, anti-IFNγ mAb, IL-23, IL-6, IL-1β, FICZ. For proliferation assays, sorted naïve CD4 T cells were labeled with 5 μM CFSE (eBioscience) prior to culture. Cells were cultured in a standard CO2 incubator, with 5% CO2, approximately 70% humidity and approximately 17% O2.
Chitrakar A., Budda S.A., Henderson J.G., Axtell R.C, & Zenewicz L.A. (2020). E3 ubiquitin ligase von Hippel-Lindau (VHL) protein promotes Th17 differentiation. Journal of immunology (Baltimore, Md. : 1950), 205(4), 1009-1023.
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9

Intradermal IL-23 and MaR1 in Mice

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Twenty microliters of PBS, either alone or containing 500 ng recombinant mouse IL-23 from eBioscience (San Diego, CA), was injected intradermally into the ears of anesthetized mice using a 30-gauge needle every other day for 16 days. Topical vehicle (ethanol) or MaR1 (100 ng in 20 μl ethanol/ear) was administrated 30 min before IL-23 injection. Thickness of both mouse ears was measured before injection on day 0 and thereafter on before injections and on day 16. The mean ear thickness of both ears was calculated for each mouse.
Saito-Sasaki N., Sawada Y., Mashima E., Yamaguchi T., Ohmori S., Yoshioka H., Haruyama S., Okada E, & Nakamura M. (2018). Maresin-1 suppresses imiquimod-induced skin inflammation by regulating IL-23 receptor expression. Scientific Reports, 8, 5522.
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