Il 18

Manufactured by MBL Life Science

Sourced in United States, Canada

IL-18 is a cytokine that is involved in the activation and regulation of the immune system. It plays a role in the production of interferon-gamma and the promotion of Th1 immune responses. IL-18 is used in research applications to study its function in the immune system.

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Lab products found in correlation

Il 12, by Thermo Fisher Scientific (5 mentions) Il 15, by R&D Systems (4 mentions) Il 12, by R&D Systems (4 mentions) Il 1β, by Thermo Fisher Scientific (4 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (4 mentions) Il 12, by MBL Life Science (3 mentions) Multiplex elisa, by Merck Group (3 mentions) Il 15, by Thermo Fisher Scientific (3 mentions) Il 1β, by BD (2 mentions) Rapamycin, by Merck Group (2 mentions) Golgistop, by BD (2 mentions) Golgiplug, by BD (2 mentions) Caspase 1 ice flourometric assay, by R&D Systems (2 mentions) Tissuelyser, by Qiagen (2 mentions) Il 21, by Thermo Fisher Scientific (2 mentions) Classical elisa for il 1β, by Thermo Fisher Scientific (2 mentions) Il 1β, by R&D Systems (1 mentions) Interleukin 2 (il 2), by Novartis (1 mentions) Dynabeads human t cell activator cd3cd28, by Thermo Fisher Scientific (1 mentions) Il 1b, by R&D Systems (1 mentions)

37 protocols using il 18

Quantification of Inflammatory Cytokines

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Plasma concentrations of IL-1β (BD Biosciences) and IL-18 (MBL International, Woburn, MA, USA) were quantified by ELISA according to the manufacturers’ instructions.
Concentrations of IL-1β in cell culture supernatants were analyzed by AlphaLISA, a bead-based immunoassay, according to the protocol provided by the manufacturer (PerkinElmer, Waltham, MA, USA). IL-18 abundance in cell culture supernatants was determined by ELISA (MBL International) as per the manufacturer’s instructions. Where indicated, cytokine abundance in cell culture supernatants was normalized to total protein concentration (determined by BCA assay; Thermo Fisher Scientific).

Rudloff I., Ung H.K., Dowling J.K., Mansell A., D’Andrea L., Ellisdon A.M., Whisstock J.C., Berger P.J., Nold-Petry C.A, & Nold M.F. (2020). Parsing the IL-37-Mediated Suppression of Inflammasome Function. Cells, 9(1), 178.

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NK Cell Activation by Cytokine Stimulation

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Purified NK cells were plated at a concentration of 2 × 10⁵ cells/well in RPMI medium supplemented with Pen/Strep/Glut and 10% FCS. The NK cells were cultured either alone or stimulated with 10 ng/ml of recombinant human IL-12 (Peprotech), IL-15 (Peprotech), IL-18 (MBL International), or the combination of IL-12, IL-15 and IL-18. In blocking experiments, the cells were incubated with Human BD Fc block (2.5 μg/ml) and 20 μg/ml anti-NKG2D (149810) or Mouse IgG1 Isotype control (11711) throughout the culture period. When indicated, the TACE inhibitor TAPI-0 (Peptides International Louisville, KY) or anti-TACE antibody were added at a concentration of 1 μM and 6 μg/ml, respectively. The cells were analyzed after 18 hours of culture. For the cell count experiments, 4 × 10⁵ cells/well supplemented with IL-12, IL-15 and IL-18 were plated by adding 20 μg/ml of anti-NKG2D or IgG1 isotype control antibodies and the live cells were counted 18 hours later.

Sharma N., Trinidad C.V., Trembath A.P, & Markiewicz M.A. (2017). NKG2D signaling between human NK cells enhances TACE-mediated TNF-α release. Journal of immunology (Baltimore, Md. : 1950), 199(8), 2865-2872.

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Western Blot and ELISA for Apoptosis

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For Western Blot analysis, cells were lysed by adding NP-40 to the media to a final concentration of 1 % and protease inhibitors. Cleared lysates were separated by 12% SDS-PAGE, transferred to PVDF membranes and probed with a rabbit anti-caspase-1 antibody generated in our laboratory. Concentrations of IL-1β(R&D Systems) and IL-18 (MBL International) were measuredby ELISA in cell-free culture supernatant.

Franchi L., Eigenbrod T., Muñoz-Planillo R., Ozkurede U., Kim Y.G., Arindam C., Gale M J.r., Silverman R.H., Colonna M., Akira S, & Núñez G. (2014). Cytosolic Double-Stranded RNA Activates the NLRP3 inflammasomevia MAVS-Induced Membrane Permeabilization and K+ Efflux. Journal of immunology (Baltimore, Md. : 1950), 193(8), 4214-4222.

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Isolation and Characterization of Ovarian Cancer Myeloid-Derived Suppressor Cells

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Primary OvCa ascites cells were harvested from bulk ascites by centrifugation. When indicated, bulk OvCa ascites cells were stimulated with combinations of IL18 (200 ng/ml; MBL International), IFNα (1000 IU/ml; Intron A, IFN-α-2b; Schering-Plough), IL12 (5 ng/ml; PeproTech), IL2 (250 IU/ml; Chiron), monoclonal antibody (mAb) to CD3 (clone OKT3; 1 μg/ml; eBioscience), and CD3/CD28 Human T cell-Activator Dynabeads (5 μl/ml; Invitrogen). NK cells were depleted from bulk OvCa ascites cells by CD56 positive magnetic selection (StemCell Technologies). MDSCs were depleted or isolated using CD11b magnetic selection (Miltenyi Biotech). This procedure has been previously shown (16 (link), 17 (link)) to be highly effective in isolating > 95% pure CD11b⁺ cells uniformly expressing the CD11b⁺CD33⁺CD14⁺HLA-DR^low/− monocytic MDSC phenotype from human OvCa ascites cells (16 (link), 17 (link)) (also CD34⁺CD80⁻CD83⁻DC-SIGN⁻ILT2-4⁺ expressing ARG1, NOS2, IDO1, IL10, COX2, and IL4Rα; please see Results and Supplemental Material). Control CD11b⁺ cells were isolated from healthy donor peripheral blood using the same method.

Wong J.L., Obermajer N., Odunsi K., Edwards R.P, & Kalinski P. (2016). Synergistic COX2 induction by IFNγ and TNFα self-limits type-1 immunity in the human tumor microenvironment. Cancer immunology research, 4(4), 303-311.

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Quantification of Secreted Cytokines

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The concentration of secreted human IL-1β and IL-18 in culture supernatants was measured by ELISA kits (IL-1β, BD Biosciences; IL-18, MBL International) according to the manufacturers’ instructions.

Weng L., Mitoma H., Tricot C., Bao M., Liu Y., Zhang Z, & Liu Y.J. (2014). The E3 ubiquitin ligase TRIM33 is essential for cytosolic RNA-induced NLRP3 inflammasome activation. Journal of immunology (Baltimore, Md. : 1950), 193(7), 3676-3682.

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Aluminum Nanoparticle Exposure on Cytokine Production

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Splenocytes were plated in 6 well plates at 500,000 cells/well, after which they were primed with LPS (10 ng/ml) for 12 h prior to being exposed to either PBS, AlO NSs (125 mg/mL) or AlO NWs (300 mg/mL) for 8 h. Culture supernatants were then collected, centrifuged for 5 min at 5,000 rpm to remove cells, and assessed for IL-1b and IL18 protein content by ELISA following the manufacturer's protocol (IL1b: R&D Systems, Minneapolis, MN, USA; IL18: MBL International, Woburg, MA, USA). Absorbance was recorded at 450 and 570 nm with an ELISA plate reader (Optima FluoStar, BMG LabTech GmbH, Ortenberg, Germany).

Manshian B.B., Poelmans J., Saini S., Pokhrel S., Grez J.J., Himmelreich U., Mädler L, & Soenen S.J. (2018). Nanoparticle-induced inflammation can increase tumor malignancy. Acta biomaterialia, 68.

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MAIT Cell Activation by E. coli-loaded Monocytes

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Healthy adult blood was obtained from the blood bank of the University Leipzig, approved by the local ethics committee (Ref. #079-15-09032015). Peripheral blood mononuclear cells were isolated from male human donors at age 20-50 by Ficoll-Paque™ density-gradient (GE Healthcare, UK) centrifugation. CD161+ TCR Vα7.2+ MAIT cells and CD14+ monocytes were obtained by positive magnetic separation using respective microbeads (Miltenyi Biotec, Germany). For RNA-Seq experiments, monocytes were loaded with fixed E. coli at a MOI of 10 for 3h, followed by intense washing in PBS to remove E. coli. MAIT cells and E. coli-loaded monocytes were incubated at a ratio of 1:1 for 16h. In some experiments, MAIT cells were stimulated with 50ng/ml IL-12 and 50ng/ml IL-18 (both from MBL International, MA, USA), 10µg/ml plate-bound anti-CD3 and 1µg/ml soluble anti-CD28 (both from Biolegend, CA, USA) or a combination of both for 16h. For TWIST1 inhibition experiments, the small molecule inhibitor harmine (Sigma-Aldrich, Germany) was used at a concentration of 20 or 40µM, DMSO served as solvent control.

Schubert K., Karkossa I., Schor J., Engelmann B., Steinheuer L.M., Bruns T., Rolle-Kampczyk U., Hackermüller J, & von Bergen M. (2021). A Multi-Omics Analysis of Mucosal-Associated-Invariant T Cells Reveals Key Drivers of Distinct Modes of Activation. Frontiers in Immunology, 12, 616967.

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Cytokine profiling of CD56+ T cells

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Cells were isolated from umbilical cord blood or iPSC T cell differentiation cultures by fluorescence-activated cell sorting on the basis of CD7 and CD56 expression. Cells were cultured in AIM V medium (no. 31035025, Thermo Fisher Scientific) containing 5% heat-inactivated FBS without or with 10 ng/mL IL-12 (Peprotech), 10 ng/mL IL-15 (R&D), and 20 ng/mL IL-18 (MBL International) in a 96 round-bottom well plate at a density of 12,000 cells/well. After 20 h the supernatants were harvested after spinning the cell suspension at 450 × g and snap frozen. Medium without cells served as baseline. Cytokine production was measured with the Bio-Plex Pro Human Cytokine 27-plex Immunoassay (Bio-Rad) according the manufacturer's protocol.

Themeli M., Chhatta A., Boersma H., Prins H.J., Cordes M., de Wilt E., Farahani A.S., Vandekerckhove B., van der Burg M., Hoeben R.C., Staal F.J, & Mikkers H.M. (2020). iPSC-Based Modeling of RAG2 Severe Combined Immunodeficiency Reveals Multiple T Cell Developmental Arrests. Stem Cell Reports, 14(2), 300-311.

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Quantifying Cytokine Levels in BMDM

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The concentrations of cytokines in the conditioned medium of BMDM cultures were determined by ELISA. IL-1β (Thermo Fisher Scientific), IL-18 (MBL International), IFN-β (PBL Assay Science) concentration was quantified using the standard ELISA development kit according to the manufacturer’s protocol.

da Costa L.S., Outlioua A., Anginot A., Akarid K, & Arnoult D. (2019). RNA viruses promote activation of the NLRP3 inflammasome through cytopathogenic effect-induced potassium efflux. Cell Death & Disease, 10(5), 346.

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Quantification of Wound Cytokines

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Mouse wounds were homogenized in cold PBS (10 μL PBS/mg wound tissue) supplemented with protease inhibitor cocktail (Sigma). Supernatants of wound homogenates or cell culture medium were used for ELISA for IL-1β, IL-6, IL-10, TGF-β1, TNF-α (eBioscience), IL-18 (MBL International), and IGF-1 (R&D Systems). When wound-conditioned medium was used as a cell culture supplement, cytokine release was measured as the difference between levels achieved in wells with cultured cells and levels in blank wells that contained identical medium composition but no cells.

Mirza R.E., Fang M.M., Weinheimer-Haus E.M., Ennis W.J, & Koh T.J. (2014). Sustained Inflammasome Activity in Macrophages Impairs Wound Healing in Type 2 Diabetic Humans and Mice. Diabetes, 63(3), 1103-1114.

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