Ril 33

Manufactured by BioLegend

Sourced in United States

RIL-33 is a laboratory instrument used for conducting cell culture and cell biology experiments. It is designed to provide a controlled environment for the cultivation and study of cells. The core function of RIL-33 is to maintain optimal temperature, humidity, and gas levels to support the growth and maintenance of cell cultures.

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Lab products found in correlation

9 protocols using ril 33

Modulation of BMMC Proliferation and Cytokine Production

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One million BMMCs per mL were cultured in triplicate with IL-3 and SCF or 2 μg/mL DNP-IgE (Sigma Aldrich). To determine the effects of TSA on proliferation and cytokine production TSA in dimethyl sulfoxide (DMSO) was added in concentrations of 10, 30,100, 300, or 500 nM for varying time points. Control wells were treated with vehicle alone. Cells were then stimulated with 200 ng/mL DNP-BSA (Sigma Aldrich) or 20 ng/mL rIL-33 (Biolegend).

Krajewski D., Kaczenski E., Rovatti J., Polukort S., Thompson C., Dollard C., Ser-Dolansky J., Schneider S.S., Kinney S.R, & Mathias C.B. (2018). Epigenetic Regulation via Altered Histone Acetylation Results in Suppression of Mast Cell Function and Mast Cell-Mediated Food Allergic Responses. Frontiers in Immunology, 9, 2414.

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Tamoxifen-Induced Mouse Genetic Manipulation

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Mice were treated with tamoxifen dissolved at 20 mg/ml in corn oil (Sigma) for Id2^CreERT2, or at 40 mg/ml in EtOH supplemented with 1 volume of Cremophor EL (Sigma C5135) and 2 volumes of PBS for a final concentration of 10 mg/ml for Arg1^RFP-CreERT2 mice. Pregnant females were treated by oral gavage with 1 mg tamoxifen and 0.5 mg progesterone (Sigma P0130) twice daily on E16, 17 and 18. Day of embryonic development was estimated by taking the day of vaginal plug observation as E0.5. Because tamoxifen treatment during pregnancy interferes with normal delivery, pups were delivered by caesarean sections on E19.5 and neonates were fostered by lactating females. Neonatal pups were injected i.p. with 1.5 mg on P10 for Id2^CreERT2, or 400 μg on P10,11 and 12 for Arg1^RFP-CreERT2 mice. Adult mice were fed a tamoxifen chow (Envigo TD.130858) for 4 weeks. IL-7Rα blockade was performed by injecting 3 doses of 200 μg InVivoMAb anti-mouse IL-7Rα (A7R34, BioXCell) i.v. into timed pregnant females on E14.5, E16.5, and E18.5. For Nur77 stimulation, mice were injected i.p. with 1 μg rIL-33 (Biolegend) 16h prior to analysis. For infections with N. brasiliensis, mice were injected subcutaneously with 500 L3-stage larvae and analyzed at the indicated timepoints post-infection.

Schneider C., Lee J., Koga S., Ricardo-Gonzalez R.R., Nussbaum J.C., Smith L.K., Villeda S.A., Liang H.E, & Locksley R.M. (2019). Tissue-resident group 2 innate lymphoid cells differentiate by layered ontogeny and in situ perinatal priming. Immunity, 50(6), 1425-1438.e5.

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Intranasal IL-33 Enhances Influenza Infection

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Mice were anesthetized via isoflurane inhalation and then injected intranasally with 0.5 μg of recombinant IL-33 (rIL-33; BioLegend, San Diego, CA, USA) in 20 μL phosphate-buffered saline (PBS) daily for 5 days prior to virus infection.
Influenza virus strain (strain A/Puerto Rico/8/1934 H1N1, PR8) was provided by A. Iwasaki (Yale University, New Haven, CT, USA). For intranasal virus infections, mice were injected intraperitoneally with ketamine (100 mg/kg) and xylazine (5.83 mg/kg) for anesthesia, followed by intranasal injections of 50 pfu PR8 in 20 μL PBS. After infection, mice were monitored for survival and body weight until body weight loss reached 75% of their initial weight.

Kim C.W., Yoo H.J., Park J.H., Oh J.E, & Lee H.K. (2019). Exogenous Interleukin-33 Contributes to Protective Immunity via Cytotoxic T-Cell Priming against Mucosal Influenza Viral Infection. Viruses, 11(9), 840.

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MCMV Infection and Viral Challenge Protocols

Cited 3 times

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Il33^−/− mice were bred in-house (38 (link)). C57BL/6 mice were purchased from Charles River Laboratories or Envigo. Sex-matched mice aged 7–9 wk were used in all experiments. Smith-strain MCMV was propagated in vivo and prepared via sorbitol gradient purification (39 (link)). Mice were infected i.p. with 3 × 10⁴ PFU of MCMV. ΔgL-SL8-MCMV was prepared as described previously (24 (link)). Mice were infected i.p. with 4 × 10⁵ PFU of ΔgL-SL8-MCMV. In some experiments, 2 μg of rIL-33 (BioLegend) was administered i.p. at the time of infection. Replication-deficient recombinant adenovirus type 5 (pAdZ5-CV5) expressing immediate-early protein 3 (rAd-IE3) was engineered and purified as described previously (40 (link)). Mice were challenged i.p. with 5 × 10⁸ PFU of rAd-IE3 or 2 × 10⁶ PFU of recombinant vaccinia virus expressing OVA (rVV-OVA) (41 (link)). All mouse experiments were performed at Cardiff University under U.K. Home Office Project License 30/2969 (London, U.K.).

McLaren J.E., Clement M., Marsden M., Miners K.L., Llewellyn-Lacey S., Grant E.J., Rubina A., Gimeno Brias S., Gostick E., Stacey M.A., Orr S.J., Stanton R.J., Ladell K., Price D.A, & Humphreys I.R. (2019). IL-33 Augments Virus-Specific Memory T Cell Inflation and Potentiates the Efficacy of an Attenuated Cytomegalovirus-Based Vaccine. The Journal of Immunology Author Choice, 202(3), 943-955.

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ILC2 Culture and Expansion Protocol

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ILC2 culture was performed as before^{41 (link)}. ILC2s isolated from each organ were cultured with complete medium (RPMI-1640 medium [Sigma–Aldrich], 10 mM HEPES [Life Technologies], 1 mM sodium pyruvate [Life Technologies], 1% MEM nonessential amino acids [Life Technologies], 0.1% 2-mercaptoethanol and L-glutamine) with 25 U/mL rIL-7Rα (BioLegend), 10 ng/mL r IL-25 (BioLegend), and 10 ng/mL r IL-33. The ILC2s were maintained at 37 °C with 5% CO₂ and passaged after 2−3 days into the same volume of medium containing cytokines. The ILC2 cell line ILC2/b6 was generously donated by Dr. Q Yang. The ILC2/b6 cells were cultured in OP9 medium (α-MEM, 20% FBS, 50 μM β-mercaptoethanol, and Pen-Step-Glutamine) supplemented with 10 ng/mL IL-2, IL-7, and IL-33 (PeproTech Inc.)^{42 (link)}.

Fujimoto M., Yokoyama M., Kiuchi M., Hosokawa H., Nakayama A., Hashimoto N., Sakuma I., Nagano H., Yamagata K., Kudo F., Manabe I., Lee E., Hatano R., Onodera A., Hirahara K., Yokote K., Miki T., Nakayama T, & Tanaka T. (2022). Liver group 2 innate lymphoid cells regulate blood glucose levels through IL-13 signaling and suppression of gluconeogenesis. Nature Communications, 13, 5408.

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Culturing Lung and Skin ILC2s

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Lin⁻CD45⁺Thy1⁺Red5⁺ ILC2s from lung and skin were FACS sorted and 2500 (lung) or 1000 (skin) cells per well were cultured in RPMI-1640 (Sigma) containing 10% FBS, 10 mM HEPES (Sigma), 100 μM non-essential amino acids (Sigma), 1 mM sodium pyruvate (Gibco), 100 μU/mL penicillin (Gibco), 100 μg/mL streptomycin (Gibco), 50 μM 2-mercaptoethanol (Gibco), 10 ng/ml rIL-7 (R&D Systems), 10 ng/ml rIL-2 (R&D Systems), and 10 ng/ml rIL-33 (BioLegend) in 96-well round bottom plates at 37 °C under 5% CO₂. After 3 days, supernatants were collected and the concentration of CXCL2 was measured by ELISA (ThermoFisher, EMCXCL2).

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IL-33 and IL-13 Effects on Mice

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Mice were administered 200 µL of sterile PBS as a control or 0.5 µg of carrier-free murine rIL-33 (BioLegend, 580506) in 200 µL of sterile PBS via daily intraperitoneal injection for 5 days, unless otherwise noted. Each mouse was used for blood glucose measurements and euthanized the day after the final injection. An independent series of mice were administered 1 µg of carrier-free murine rIL-13 (BioLegend, 575906) in a single injection and then subjected to PTT analysis the next day. Fifty micrograms of control IgG (BioLegend, 400192) or anti-mouse IL-13 neutralizing antibody (InvivoGen, mabg-mil13-5) was administered via daily intraperitoneal injection for 4 days.

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Murine LCMV-induced Hepatitis Model

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C57BL/6 (B6) mice from the Jackson Laboratory and IL-33^−/− mice of the B6 background [48 (link)] were bred and maintained under specific pathogen-free conditions in the UTMB animal care facility and used at 7–10 wk of age. All procedures were approved by UTMB’s Institutional Animal Care and Use Committee and performed according to the NIH Guidelines. In all experiments, LCMV strain Clone 13 (Cl 13, a gift from Dr. Maria Salvato at the University of Maryland) was used, and mice were i.v. injected with 2 × 10⁶ PFU of viruses to induce hepatitis. Some infected mice were i.p. injected with rIL-33 (1 μg/mouse, Biolegend, San Diego, CA) or PBS, at indicated time points.

Liang Y., Jie Z., Hou L., Yi P., Wang W., Kwota Z., Salvato M., de Waal Malefyt R., Soong L, & Sun J. (2015). IL-33 promotes innate IFN-γ production and modulates dendritic cell response in LCMV-induced hepatitis in mice. European journal of immunology, 45(11), 3052-3063.

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Glucose and Insulin Tolerance Assays

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For glucose tolerance tests, mice were fasted overnight before the i.p. injection of dextrose (2 mg/gr). For insulin tolerance tests, mice were fasted 6 hours before i.p. insulin administration (0.5 U/kg). Tail vein blood glucose levels were measured at indicated time points using glucose meters (Bayer). When indicated, mice were i.p. injected every other day with either 500 ng murine rIL-33 (BioLegend; Cat #580502) or PBS to obtain a total of three doses. Mice were analyzed 24 hours after the last injection.

Burganova G., Schonblum A., Sakhneny L., Epshtein A., Wald T., Tzaig M, & Landsman L. (2023). Pericytes modulate islet immune cells and insulin secretion through Interleukin-33 production in mice. Frontiers in Endocrinology, 14, 1142988.

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