Il 18 elisa kit

Manufactured by R&D Systems

Sourced in United States

The IL-18 ELISA kit is a quantitative sandwich enzyme immunoassay designed for the measurement of interleukin-18 (IL-18) levels in cell culture supernatants, serum, and plasma samples. The kit utilizes a monoclonal antibody specific for IL-18 that is pre-coated onto a microplate. Samples and standards are pipetted into the wells, and any IL-18 present is bound by the immobilized antibody. After washing, an enzyme-linked polyclonal antibody specific for IL-18 is added to the wells. Following a wash to remove any unbound antibody-enzyme reagent, a substrate solution is added and color develops in proportion to the amount of IL-18 bound. The intensity of the color is measured at a specific wavelength.

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Revertaid first strand cdna synthesis kit, by Thermo Fisher Scientific (2 mentions) Il 1β elisa kit, by R&D Systems (1 mentions) Glutamine, by Thermo Fisher Scientific (1 mentions) Bay11 7082, by Merck Group (1 mentions) Polyvinylidene fluoride (pvdf), by Merck Group (1 mentions) Pf06650833, by Merck Group (1 mentions) Zyvad ome fmk, by Enzo Life Sciences (1 mentions) Fetal bovine serum (fbs), by MP Biomedicals (1 mentions) Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions) Opti mem, by Thermo Fisher Scientific (1 mentions) Rpmi 1640 medium, by Cytiva (1 mentions) Protease inhibitor cocktail, by Merck Group (1 mentions) Phenylmethylsulfonyl fluoride, by Merck Group (1 mentions) Pro prep solution, by iNtRON Biotechnology (1 mentions) Il 1β elisa kit, by BD (1 mentions) Trizol reagent, by Thermo Fisher Scientific (1 mentions) Sybr green master mix, by Qiagen (1 mentions)

Close spelling variants of this product

IL-1β and IL-18 ELISA kits Mouse IL-18 DuoSet ELISA kit Mouse IL-18/IL-1F4 ELISA kit Human IL-18 BPaQuantikine ELISA Kit Human Total IL-18/IL-1F4 Quantikine ELISA Kit Human IL-18 ELISA kit Mouse IL-18 ELISA Kit IL-18 ELISAs

7 protocols using il 18 elisa kit

YKL-40 Stimulation of Osteoblast IL-18

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Osteoblasts were pretreated with pharmacological inhibitors or transfected with siRNA, then stimulated with YKL-40 for 24 h. CM was collected and stored at −80 °C. IL-18 expression in CM was examined using the IL-18 ELISA kit (R&D Systems, Minneapolis, MN, USA), according to the manufacturer’s procedure.

Li T.M., Liu S.C., Huang Y.H., Huang C.C., Hsu C.J., Tsai C.H., Wang S.W, & Tang C.H. (2017). YKL-40-Induced Inhibition of miR-590-3p Promotes Interleukin-18 Expression and Angiogenesis of Endothelial Progenitor Cells. International Journal of Molecular Sciences, 18(5), 920.

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Quantifying Inflammatory Cytokines in Sera

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The concentrations of inflammatory cytokines IL-1β, IL-6, IL-8, IL-10, and TNFα were performed using a commercially available ELISA obtained from PeproTech (PeproTech, Neuilly-sur-Seine, France). IL-18 ELISA kit was purchased from R&D, (France). Levels of cytokines were quantified in the patient sera according to the manufacturer’s protocols. The levels of the cytokines measured were in a linear range of a standard curve as previously described [30 (link)].

Reynaud D., Abi Nahed R., Lemaitre N., Bolze P.A., Traboulsi W., Sergent F., Battail C., Filhol O., Sapin V., Boufettal H., Hoffmann P., Aboussaouira T., Murthi P., Slim R., Benharouga M, & Alfaidy N. (2021). NLRP7 Promotes Choriocarcinoma Growth and Progression through the Establishment of an Immunosuppressive Microenvironment. Cancers, 13(12), 2999.

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Evaluating the Role of USP5 and TXNIP in LPS-Induced Inflammatory Response

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Huh7 or HepG2 cells were seeded into 6-well plates at a density of 100,000 cells per well for 12 hours and were divided into 6 groups: MOCK (phosphate buffer solution), LPS, LPS+shNC, LPS+shUSP5, LPS+shUSP5+Vector, and LPS+shUSP5+TXNIP. After transfection with 5-µg plasmid (shNC, shUSP5, Vector, or TXNIP) for 24 hours, 70% confluence of cells in 6-well plates were incubated with 1000 ng/mL LPS for 24 hours. Subsequently, the supernatant was collected for IL-1β and IL-18 detection using IL-1β ELISA kit (CAT#: RLB00, R&D Systems, Hinnerup, Denmark) and IL-18 ELISA kit (CAT#: DY3144-05, R&D Systems) as described.²⁰ The sample concentrations were calculated according to the standard curve obtained.

Shi S., Pan X., Chen M., Zhang L., Zhang S., Wang X., Shi S., Chen Z., Lin W, & Jiang Y. (2023). USP5 promotes lipopolysaccharide-induced apoptosis and inflammatory response by stabilizing the TXNIP protein. Hepatology Communications, 7(8), e0193.

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NLRP3 Inflammasome Activation Pathway Protocol

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Roswell Park Memorial Institute (RPMI) 1640 medium was purchased from Hyclone Laboratories (South Logan, UT, USA). Fetal bovine serum (FBS) was obtained from MP Biomedicals (Santa Ana, CA, USA). Opti-MEM, glutamine and penicillin/streptomycin were the products of Gibco (Grand island, NY, USA). Isoliquiritigenin (1), phorbol 12-myristate 13-acetate (PMA), phenylmethylsulfonyl fluoride (PMSF), polyvinylidene fluoride (PVDF), PF 06650833 (IRAK4 inhibitor), Bay 11-7082 (NF-κB inhibitor) and protease inhibitor cocktail were purchased from Sigma-Aldrich (St. Louis, MO, USA). Suberic acid bis (3-sulfo-N-hydroxysuccinimide ester) sodium salt (BS³) was bought from BioVision (Milpitas, CA, USA). Lactate dehydrogenase (LDH) cytotoxicity WST assay kit and Z-YVAD(Ome)-FMK (caspase-1 inhibitor) were obtained from Enzo Life Sciences (Farmingdale, NY, USA). Pro-prep solution was obtained from iNtRON Biotechnology (Seongnam, Korea). All primary antibodies relating NLRP3 inflammasome and signaling pathway and secondary antibodies were bought form Cell Signaling Technology (Minneapolis, MN, USA). Primary anti-β-actin antibody was obtained from Bethyl Laboratories (Montgomery, TX, USA). IL-1β ELISA kit was purchased from BD Biosciences (San Jose, CA, USA). IL-18 ELISA kit was bought from R&D Systems (Minneapolis, MN, USA).

Choi H.R., Lim H., Lee J.H., Park H, & Kim H.P. (2021). Interruption of Helicobacter pylori-Induced NLRP3 Inflammasome Activation by Chalcone Derivatives. Biomolecules & Therapeutics, 29(4), 410-418.

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Quantifying Cytokine Levels and Gene Expression

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Mouse IL-1β, IL-18 ELISA kits were purchased from R&D Systems. The levels of cytokines were determined according to the manufacturer’s instructions. IL-1β procedure was modified with orbital shaking to decrease the lower detection limit of the kit to 2.35 pg/mL as previously described^{35 (link)}. Total RNA was isolated from mice tissue using TRIZOL Reagent (Invitrogen, Carlsbad, CA) following the manufacturer’s protocol. 1ug RNA was reversely transcribed to complementary DNA(cDNA) using Revert Aid First Strand cDNA Synthesis Kit (Thermo Fisher Scientific, San Jose, CA). For Real-time PCR(RT-PCR), equal amounts of cDNA were added to SYBR Green master mix (Qiagen, Valencia,CA) to detect target transcripts using Roche light cycler 480. Data for relative expression were presented using the 2ΔΔCt method.

Kearns A.C., Liu F., Dai S., Robinson J.A., Kiernan E., Tesfaye Cheru L., Peng X., Gordon J., Morgello S., Abuova A., Lo J., Zanni M.V., Grinspoon S., Burdo T.H, & Qin X. (2019). Caspase-1 activation is related with HIV-associated atherosclerosis in a HIV transgenic mouse model and HIV patient cohort. Arteriosclerosis, thrombosis, and vascular biology, 39(9), 1762-1775.

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Cytokine Quantification by ELISA

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We centrifuged media in which cells had been cultured for 72 h at 12,000 rpm at 4°C for 10 min to remove debris. The supernatants were collected for enzyme‐linked immunosorbent assays (ELISAs) using human IL‐1β and IL‐18 ELISA kits (R&D Systems) according to the manufacturer's instruction.

Liu X., Wang C., Pang L., Pan L, & Zhang Q. (2022). Combination of resolvin E1 and lipoxin A4 promotes the resolution of pulpitis by inhibiting NF‐κB activation through upregulating sirtuin 7 in dental pulp fibroblasts. Cell Proliferation, 55(5), e13227.

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Quantifying IL-1β and IL-18 Levels

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The levels of IL-1β and IL-18 were measured using IL-1β ELISA kits (MLB00C; R&D Systems, Minneapolis, MN) and IL-18 ELISA kits (7625, R&D Systems) [17 (link)]. Each assay was performed according to the manufacturer’s instructions. The optical density was determined at 450 nm using an absorption spectrophotometer (Finstruments Multiskan Model 347). Optical density values used for quantitative analysis were averages of five measurements.

Zhu J., Chen L., Qi Y., Feng J., Zhu L., Bai Y, & Wu H. (2018). Protective effects of Erigeron breviscapus Hand.– Mazz. (EBHM) extract in retinal neurodegeneration models. Molecular Vision, 24, 315-325.

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