Mouse il 1β elisa kit

Manufactured by BD

Sourced in United States

The Mouse IL-1β ELISA kit is a sandwich enzyme-linked immunosorbent assay (ELISA) designed for the quantitative measurement of mouse interleukin-1 beta (IL-1β) in various biological samples, such as cell culture supernatants, tissue homogenates, and serum. It utilizes a specific antibody coated on a microplate to capture the target analyte, and a detection antibody labeled with an enzyme for colorimetric signal development and quantification.

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Lab products found in correlation

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10 protocols using mouse il 1β elisa kit

Cytokine Profiling in MAYV Infection

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For in vitro cytokine determination, BMDMs were seeded overnight at a density of 2 × 10⁵ cells/well in 48-well plates and prestimulated with 300 ng/ml of PAM(3)CSK(4) (Invivogen) for 4 h, and subsequently infected with MAYV. The cytokines in the supernatants were assayed using a mouse IL-1β ELISA kit (BD Biosciences) according to the manufacturer’s instructions. For in vivo cytokine determination, footpads were processed and the supernatants from total homogenates were obtained. The levels of IL-1β and IL-18 in the homogenates were detected using a mouse IL-1β ELISA kit (BD Biosciences) and a mouse IL-18 ELISA kit (RD-MBL), respectively, according to the manufacturer’s instructions. Detection of IL-1β, IL-18 and Caspase 1 (p20 subunit) in human serum samples was accomplished by using human IL-1β ELISA kit II (BD Biosciences), human total IL-18 (RD) and human caspase-1 ELISA kit (RD) respectively.

de Castro-Jorge L.A., de Carvalho R.V., Klein T.M., Hiroki C.H., Lopes A.H., Guimarães R.M., Fumagalli M.J., Floriano V.G., Agostinho M.R., Slhessarenko R.D., Ramalho F.S., Cunha T.M., Cunha F.Q., da Fonseca B.A, & Zamboni D.S. (2019). The NLRP3 inflammasome is involved with the pathogenesis of Mayaro virus. PLoS Pathogens, 15(9), e1007934.

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Quantifying Cytokine Levels in Spleen

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Splenic concentrations of TNF-α, IL-1β and IL-10 were measured by ELISA using commercially available kits, including a mouse TNF-α ELISA kit (cat. no. 558534, BD Biosciences), a mouse IL-1β ELISA kit (cat. no. MBS824757) and a mouse IL-10 ELISA kit (cat. no. MBS8244590; both MyBioSource, Inc., San Diego, CA, USA), respectively, as previously described (18 (link)). Tissue samples (10–15 mg) were homogenized in a tissue grinder containing 1 ml lysis buffer (PBS containing 2 mM phenylmethylsulfonyl fluoride and 1 mg/ml aprotinin, leupeptin, and pepstatin A, all obtained from Sigma-Aldrich; Merck KGaA), as described by Clark et al (21 (link)). Analysis was performed with 100 ml standard (diluted in lysis buffer) or 10, 50 or 100 ml tissue homogenate. Each sample was run in duplicate and a portion of the sample was analyzed for protein. Data were expressed as pg/mg of protein. For each assay, a standard curve was generated and based on replicates of the measured absorbance, demonstrated an average coefficient of variance of <10%.

Kim C.G., Lee J.E., Jeong D.G., Lee Y.H., Park S.I., Lee D.G., Han C.H., Kang S.J., Song C.H., Choi S.H., Lee Y.J, & Ku S.K. (2017). Bathing effects of east saline groundwater concentrates on allergic (atopic) dermatitis-like skin lesions induced by 2,4-dinitrochlorobenzene in hairless mice. Experimental and Therapeutic Medicine, 13(6), 3448-3466.

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In Vitro Cytokine and Cell Death Assays

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For in vitro cytokine determination and LDH assay, BMDMs were seeded at a density of 2 × 10⁵ cells/well in 48-well plates and pre-stimulated with 500 ng ml⁻¹ of LPS (tlrl-peklps; InvivoGen) for 3h, and subsequently infected with CHIKV. The cytokines in the supernatants were measured using a mouse IL-1β ELISA kit (BD Biosciences) according to the manufacturer’s instructions. LDH measurements were performed with CytoTox 96 Non-Radioactive Cytotoxicity Assay (Promega) following the manufacturer’s instructions. The positive control for complete cell lysis and normalization was 9% Triton X-100 (Fisher Scientific); it was incubated with cells for 15 min. The percentage of LDH release was calculated as (mean OD value of sample / mean OD value of Triton X-100 control sample) × 100 and is shown in the figures as the percentage of cell death compared to TritonX-100 (%).

Soares-Schanoski A., Baptista Cruz N., de Castro-Jorge L.A., de Carvalho R.V., dos Santos C.A., da Rós N., Oliveira Ú., Costa D.D., dos Santos C.L., Cunha M.D., Oliveira M.L., Alves J.C., Océa R.A., Ribeiro D.R., Gonçalves A.N., Gonzalez-Dias P., Suhrbier A., Zanotto P.M., de Azevedo I.J., Zamboni D.S., Almeida R.P., Ho P.L., Kalil J., Nishiyama MY J.u.n.i.o.r, & Nakaya H.I. (2019). Systems analysis of subjects acutely infected with the Chikungunya virus. PLoS Pathogens, 15(6), e1007880.

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Inflammasome Activation Assay Protocol

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LPS (O111:B4, L2630), ATP (A7699), disuccinimidyl suberate (S1885), nigericin (N7143), and phorbol myristate acetate (PMA, P1585) were from Sigma-Aldrich; alum (77161) and the c-Myc tag (MA1-980) were from Thermo Fisher Scientific; and poly(dA:dT) was from InvivoGen. The following antibodies were used: anti-Flag (F1804) (Sigma); anti-mouse caspase-1 p20 (AG-20B-0042) and anti-mouse NLRP3 (AG-20B-0014) (AdipoGen); anti-human caspase-1 (ab108362) and anti-BRCC36 (ab108411) (Abcam); anti-mouse IL-1β (AF-401-NA) (R&D); anti-HA-tag (3724) (Cell Signaling Technology); and anti-VDR (sc-13133), anti-ASC (sc-514414), and anti-β-actin (sc-47778) (Santa Cruz). The Mouse TNF ELISA Kit (558534), Mouse IL-1β ELISA Kit (559603), and TMB Substrate Reagent Set (555214) were from BD Biosciences. The mouse IL-18 ELISA set (EMC06) was from ExCell Biotech (Table 1).

Rao Z., Chen X., Wu J., Xiao M., Zhang J., Wang B., Fang L., Zhang H., Wang X., Yang S, & Chen Y. (2019). Vitamin D Receptor Inhibits NLRP3 Activation by Impeding Its BRCC3-Mediated Deubiquitination. Frontiers in Immunology, 10, 2783.

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Inflammasome Activation Assay

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The following reagents were used: ultra-pure LPS, nigericin, poly(deoxyadenylic-deoxythymidylic) acid [poly(dA:dT)]/lyoVec, BAPTA-AM, calcium-free DMEM, disuccinimidyl suberate, Lipofectamine 2000 (all Invivogen), calcium chloride solution, thapsigargin, ATP (Sigma-Aldrich), CI-amidine (Millipore), BB-CI-amidine and GSK484 (Cayman Chemicals), AFM 30a (P. Thompson, University of Massachusetts Medical School, Worcester, MA) (21 (link)), iScript cDNA kit and iQ SYBR green supermix (BioRad), TRIzol (Ambion, Life technologies), fixation/permeabilization solution, mouse IL-1β ELISA kit (BD Bioscience), propidium iodide (ImmunoChemistry Technologies) and mouse TNF- alpha ELISA (eBioscience). Abs for confocal microscopy were ASC (AdipoGen), peptidyl-citrulline, clone F95 (Millipore), anti-vimentin (Abcam), goat anti-rabbit IgG AF647, goat anti-mouse IgM AF488 (Thermo Fisher Scientific), and goat anti-mouse IgM isotype control (SouthernBiotech). DAPI was from Molecular Probes.

Mishra N., Schwerdtner L., Sams K., Mondal S., Ahmad F., Schmidt R.E., Coonrod S.A., Thompson P.R., Lerch M.M, & Bossaller L. (2019). Protein Arginine Deiminase 2 and 4 regulate NLRP3 inflammasome- dependent IL-1β maturation and ASC speck formation in macrophages. Journal of immunology (Baltimore, Md. : 1950), 203(4), 795-800.

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Mouse IL-1β ELISA Protocol

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IL-1β in culture supernatants was assessed with a mouse IL-1β ELISA Kit (BD Biosciences, San Diego, CA, USA) according to the manufacturer's instructions. Culture medium was centrifuged at 500 g for 5 min, the supernatant was collected, and levels of IL-1β measured (39 (link)).

Lan T., Tao A., Xu X., Kvietys P, & Rui T. (2020). Peroxynitrite/PKR Axis Modulates the NLRP3 Inflammasome of Cardiac Fibroblasts. Frontiers in Immunology, 11, 558712.

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Measurement of Inflammatory Cytokines

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The concentrations of interleukin (IL)-6, tumor necrosis factor (TNF), and IL-1β in the supernatant of homogenized tongue, and serum samples were measured using a mouse IL-6 ELISA kit, mouse TNF ELISA kit, and mouse IL-1β ELISA kit (all from BD Bioscience, NJ, USA), respectively, according to the manufacturer’s instructions. The cytokine levels were estimated by interpolation from a standard curve generated using an ELISA reader (Molecular Devices) at a wavelength of 450 nm.

Lee H.R., Yoo N., Kim J.H., Sohn K.Y., Kim H.J., Kim M.H., Han M.Y., Yoon S.Y, & Kim J.W. (2016). The Therapeutic Effect of PLAG against Oral Mucositis in Hamster and Mouse Model. Frontiers in Oncology, 6, 209.

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Cytokine and Metabolic Profiling in Mice

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Mouse IL-1β ELISA kit was from BD Biosciences (559603).
Mouse IL-18 ELISA kit was from eBioscience (BMS618/3). ELISA assays were performed according manufacturer's instructions. Mouse cytokines, lipoprotein profiles, and cholesterol levels in serum were measured by core biochemical assay laboratory of Cambridge University Hospitals as described. 6 (link)

Clement M., Chen X., Chenoweth H.L., Teng Z., Thome S., Newland S.A., Harrison J., Yu X., Finigan A.J., Mallat Z, & Li X. (2019). MARK4 (Microtubule Affinity-Regulating Kinase 4)-Dependent Inflammasome Activation Promotes Atherosclerosis-Brief Report. Arteriosclerosis, thrombosis, and vascular biology, 39(8).

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Cytokine Quantification by ELISA and MSD

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Mouse IL-1β ELISA kits were obtained from BD Biosciences (catalog 559603) and Thermo Fisher Scientific (catalog 88-7013-88), and mouse TNF-α was obtained from R&D Systems (catalog DY410). Mouse IL-6 was obtained from Thermo Fisher Scientific (catalog 88-7064-88), and mouse IL-12 p70 was obtained from Thermo Fisher Scientific (catalog 88-7121-88). ELISAs were performed according to the manufacturer’s instructions. Meso Scale Discovery (MSD) assays were performed by the core biochemical assay laboratory of Cambridge University Hospitals. The detection limit of mouse IL-1β by MSD was 1.2 pg/mL.

Baldrighi M., Doreth C., Li Y., Zhao X., Warner E., Chenoweth H., Kishore K., Umrania Y., Minde D.P., Thome S., Yu X., Lu Y., Knapton A., Harrison J., Clarke M., Latz E., de Cárcer G., Malumbres M., Ryffel B., Bryant C., Liu J., Lilley K.S., Mallat Z, & Li X. (2023). PLK1 inhibition dampens NLRP3 inflammasome–elicited response in inflammatory disease models. The Journal of Clinical Investigation, 133(21), e162129.

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Cytokine Response to K+ Depletion

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Cells were plated on non-treated 6-well plates at 300,000 cells/well and allowed for growth overnight. On day 2, cells were vehicle-treated or primed with LPS (100 ng/ml) for 4 h, followed by incubation with regular culture medium containing 5 mM K⁺ or K⁺-free media in which K⁺ was replaced by Na⁺ for 2 h. Supernatant was collected and clarified and cytokine levels were determined with mouse IL-1β ELISA kits (BD Biosciences) following the manufacturer's protocol.

Zhang T., Zhao J., Liu T., Cheng W., Wang Y., Ding S, & Wang R. (2022). A novel mechanism for NLRP3 inflammasome activation. Metabolism Open, 13, 100166.

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