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Mouse duoset elisa kit

Manufactured by R&D Systems
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Mouse DuoSet ELISA kits are laboratory equipment designed for the quantitative measurement of specific mouse proteins using the enzyme-linked immunosorbent assay (ELISA) technique. They provide the necessary components to develop an ELISA for the target analyte.

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Lab products found in correlation

Protease inhibitor cocktail, by Merck Group (2 mentions) Epr3099, by Abcam (2 mentions) Anti β actin ac 15, by Merck Group (2 mentions) Mouse elisa ready set go kit, by Thermo Fisher Scientific (2 mentions) Pierce bca protein assay kit, by Thermo Fisher Scientific (1 mentions) Duoset ancillary reagent kit 2, by R&D Systems (1 mentions) Bca assay, by Thermo Fisher Scientific (1 mentions) Synergy h1 microplate reader, by Agilent Technologies (1 mentions) Duoset elisa ancillary reagent kit 2, by R&D Systems (1 mentions) Mouse 5 plex proinflammatory panel 1 kit, by Mesoscale (1 mentions) Mouse creatine kinase mb elisa kit, by Novus Biologicals (1 mentions) Anti f4 80, by BioLegend (1 mentions) Anti il 1β, by BD (1 mentions) Anti nlrp3, by R&D Systems (1 mentions) Softmax pro software, by Molecular Devices (1 mentions) Ecl detection kit, by PerkinElmer (1 mentions) Colorimetric assay kit, by Abcam (1 mentions) Bio plex pro mouse cytokine 23 plex assay, by Bio-Rad (1 mentions) Complete mini protease inhibitor cocktail, by Roche (1 mentions) Bradykinin, by Enzo Life Sciences (1 mentions)

Close spelling variants of this product

DuoSet ELISA kits (1131 citations) DuoSet ELISA (635 citations) ELISAs (144 citations) Mouse ELISA kits (102 citations) DuoSets (49 citations) Mouse IFN-γ DuoSet ELISA kit (16 citations) Mouse TNF-alpha DuoSet ELISA Kit (7 citations) Mouse CXCL1/KC DuoSet ELISA kit (7 citations) Mouse IL-1β DuoSet ELISA kit (7 citations) Mouse IL-1β/IL-1F2 DuoSet ELISA kit (4 citations)

44 protocols using mouse duoset elisa kit

1

Cytokine Profiling of Bone Marrow

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The BM content was fractioned into hematopoietic and supernatant portions by centrifugation at 1600 rpm. The supernatant fraction was re-suspended in 500 µl of 1 X PBS (supplemented with Protease Cocktail Inhibitor, Sigma). Total protein content was estimated by Pierce BCA Protein Assay Kit (ThermoFisher Scientific). Subsequently, mouse TNFα, Il1β, Il6, Cxcl1, and Ccl2 ELISAs were carried out according to the manufacturers' instructions. All Mouse DuoSet ELISA kits were purchased from R&D systems. The individual protein contents were finally normalised to total protein content.
Boroumand P., Prescott D.C., Mukherjee T., Bilan P.J., Wong M., Shen J., Tattoli I., Zhou Y., Li A., Sivasubramaniyam T., Shi N., Zhu L.Y., Liu Z., Robbins C., Philpott D.J., Girardin S.E, & Klip A. (2022). Bone marrow adipocytes drive the development of tissue invasive Ly6Chigh monocytes during obesity. eLife, 11, e65553.
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2

Plasma Cytokine Quantification via ELISA

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Mouse TNF-α, IL-6, and IL-1α cytokines were measured in mouse plasma using mouse DuoSet ELISA kits (DY410-05, DY406-05, DY400-05, R&D Systems) combined with DuoSet Ancillary reagent kit 2 (DY008, R&D Systems) according to the manufacturer’s instructions. Briefly, capture antibodies were diluted to working concentration with ELISA plate-coating buffer indicated in the certificate of analysis and incubated overnight at room temperature. Plasma samples were diluted at a 1:3 ratio in the reagent diluent and applied for further assay.
Oh T.S., Zabalawi M., Jain S., Long D., Stacpoole P.W., McCall C.E, & Quinn M.A. (2022). Dichloroacetate improves systemic energy balance and feeding behavior during sepsis. JCI Insight, 7(12), e153944.
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3

Chemokine Multiplex Assay in Spinal Cord

Cited 1 time
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Chemokine multiplex assay was performed by Quansys Biosciences. Briefly, lumbar spinal cords were collected following previously published methods (Cao et al., 2009 (link)). First, tissues were sonicated in PBS with1:100 dilution of protease inhibitor cocktail (Sigma). Then supernatants were collected following centrifugation (10000 × g at 4°C for 15 min) and stored at −80°C until assay. Prior to data collection, technicians at Quansys performed extensive pilot analyses to determine the assay buffer and sample dilutions. Total of 8 chemokines were included in the manufacturer’s pre-designed multiplex assay: CXCL1 (KC), CCL1 (TCA-3), CCL2 (MCP-1), CCL3 (MIP-1α), CCL5 (RANTES), CCL11 (Eotaxin), CCL17 (TARC), and CCL22 (MDC). In follow-up experiments, levels of CCL5 were determined via the enzyme-linked immunosorbent assay (ELISA) using a mouse DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA) following the manufacturers’ protocols. The standard series ranged from 0 – 1000 pg/ml for CCL5. Tissue levels of chemokines were normalized to the total protein levels of each sample, as determined by BCA assays (ThermoFisher Scientific, Waltham, MA).
Malon J.T, & Cao L. (2016). Calcitonin gene-related peptide contributes to peripheral nerve injury-induced mechanical hypersensitivity through CCL5 and p38 pathways. Journal of neuroimmunology, 297, 68-75.
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4

Cytokine and Chemokine Profiling in Mice

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The levels of TNF, IL-1β, IL-6, IL-10, keratinocyte chemoattractant (KC/CXCL1), monocyte chemoattractant protein-1 (MCP-1/CCL2), regulated on activation, normal T cell expressed and secreted (RANTES/CCL5), and eotaxin-1/CCL11 were measured by Mouse DuoSet ELISA kits (R&D Systems, Minneapolis, MN, USA), and the level of macrophage inflammatory protein-2 (MIP-2/CXCL2) was determined using the Mouse MIP2 Matched Antibody Pair Kit (Abcam, Cambridge, UK). The ELISA kits from both manufacturers were used in combination with the DuoSet ELISA Ancillary Reagent Kit 2 (R&D Systems). All assays were performed according to the manufacturers’ instructions with only minor modifications, and optical densities at 450 and 540 nm were measured on the Synergy H1 microplate reader (BioTek Instruments, Winooski, VT, USA).
Chlastáková A., Kaščáková B., Kotál J., Langhansová H., Kotsyfakis M., Kutá Smatanová I., Tirloni L, & Chmelař J. (2023). Iripin-1, a new anti-inflammatory tick serpin, inhibits leukocyte recruitment in vivo while altering the levels of chemokines and adhesion molecules. Frontiers in Immunology, 14, 1116324.
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5

Cytokine Quantification in Cell Media

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We measured the concentrations of MCP-1/CCL2 and MIP-1β/CCL4 inflammatory cytokines in the collected cell media of each experimental group using MCP-1 (DY479) and MIP-1β (DY451) Mouse DuoSet ELISA kits (R&D Systems) according to the manufacturer’s instructions.
Valentine M.S., Link P.A., Herbert J.A., Kamga Gninzeko F.J., Schneck M.B., Shankar K., Nkwocha J., Reynolds A.M, & Heise R.L. (2018). Inflammation and Monocyte Recruitment Due to Aging and Mechanical Stretch in Alveolar Epithelium are Inhibited by the Molecular Chaperone 4-Phenylbutyrate. Cellular and Molecular Bioengineering, 11(6), 495-508.
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6

Quantifying Inflammatory Markers in Mice

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IFNβ and IL-6 concentrations in organ homogenates were analyzed using mouse DuoSet ELISA kits (R&D Systems). Quantification of serum IL-6, IL-8, IL-1β, and TNFα was performed using a mouse V-PLEX Proinflammatory Panel 1 kit (Meso Scale Diagnostics) and was analyzed by the OSU Center for Clinical and Translational Science. Creatine kinase quantification was performed using the Mouse Creatine Kinase MB ELISA Kit (Novus Biologicals).
Kenney A.D., Aron S.L., Gilbert C., Kumar N., Chen P., Eddy A., Zhang L., Zani A., Vargas-Maldonado N., Speaks S., Kawahara J., Denz P.J., Dorn L., Accornero F., Ma J., Zhu H., Rajaram M.V., Cai C., Langlois R.A, & Yount J.S. (2022). Influenza virus replication in cardiomyocytes drives heart dysfunction and fibrosis. Science Advances, 8(19), eabm5371.
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7

Measuring CCL2, IL-1α, and TNFα in BALF

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Mouse DuoSet® ELISA kits (R&D Systems, Minneapolis, MN, United States) were used to measure levels of CCL2 (DY479), IL-1α (DY400), and TNFα (DY410) in the BALF and/or in macrophage supernatants following manufacturer’s instructions.
Milad N., Pineault M., Tremblay F., Routhier J., Lechasseur A., Beaulieu M.J., Aubin S, & Morissette M.C. (2022). Smoking status impacts treatment efficacy in smoke-induced lung inflammation: A pre-clinical study. Frontiers in Pharmacology, 13, 971238.
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8

Macrophage Responses to N. aromaticivorans

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Macrophages were harvested from the peritoneal cavity of WT and Lgals3−/− untreated mice using peritoneal lavage. Complete Dulbecco's modified Eagle's medium (6 ml) supplemented with FBS (10%), glutamine (2 mM), 100 U/ml penicillin, and 100 mg/ml streptomycin was injected into the peritoneal cavity and the medium containing cells was retrieved. The extracted cells were depleted of red blood cells using red blood cell lysis buffer containing 8.3 g/l ammonium chloride in 10 mM Tris-HCl, pH 7.5 and washed by centrifugation.
Cells were stimulated with N. aromaticivorans for 24 h (cell/bacteria ratio 1:10) at 37°C in a 5% CO2 incubator. Where indicated, cells were preincubated with the caspase-1 inhibitor Z-YVAD-FMK (10 μmol/L; Bachem AG, Bubendorf, Switzerland). After incubation, the cell supernatants were collected and cells were labeled with anti-F4/80 (BioLegend), anti-IL-1β (BD Pharmingen), and anti-NLRP3 (R&D systems) fluorochrome-conjugated monoclonal antibodies or isotype matched controls for flow cytometry. The levels of IL-1β and IL-6 in cell supernatants were determined using mouse Duoset ELISA kits (R&D Systems).
Arsenijevic A., Milovanovic J., Stojanovic B., Djordjevic D., Stanojevic I., Jankovic N., Vojvodic D., Arsenijevic N., Lukic M.L, & Milovanovic M. (2019). Gal-3 Deficiency Suppresses Novosphyngobium aromaticivorans Inflammasome Activation and IL-17 Driven Autoimmune Cholangitis in Mice. Frontiers in Immunology, 10, 1309.
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9

Splenocyte Cytokine Response to T. cruzi Antigen

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Splenocytes were harvested from wild-type mice inoculated with WT or DKO mutant parasites. The cells were plated at a concentration of 4 × 106 cells/ml in RPMI 1640 medium supplemented with 10% FBS, 100 μg/ml streptomycin, and 100 U/ml penicillin, and 1% HEPES. The non-adherent cells were removed and adherent cells (consist of mostly macrophages) were tested separately from total splenocytes. After stimulation of cells with 20 μg/ml T. cruzi freeze-thaw whole-cell antigen for 72 h, supernatants were collected, and the production of cytokines were measured by sandwich enzyme-linked immunosorbent assay (ELISA). Serum from inoculated mice were also tested for cytokine levels. Cytokines were analyzed by using Mouse Duoset ELISA kits (R&D Systems, Minneapolis, MN, USA) as per manufacturer’s instructions. Cytokine concentrations were measured at 450 nm with wavelength correction at 570 nm using SpectraMax microplate reader and data were analyzed by Softmax Pro Software (Molecular Devices LLC, Sunnyvale, CA, USA). The spleens for 4–5 animals per group were assayed.
Jha B.K., Varikuti S., Verma C., Shivahare R., Bishop N., Dos Santos G.P., McDonald J., Sur A., Myler P.J., Schenkman S., Satoskar A.R, & McGwire B.S. (2023). Immunization with a Trypanosoma cruzi cyclophilin-19 deletion mutant protects against acute Chagas disease in mice. NPJ Vaccines, 8, 63.
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10

BALF Cytokine and Chemokine Quantification

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Chemokine and cytokine levels including monocyte chemoattractant protein-1 (MCP-1), chemokine ligand 1 (CXCL1), macrophage inflammatory protein 2 (MIP-2), and interleukin-6 (IL-6) were quantified from collected BALF utilizing the Mouse DuoSet ELISA kits (R&D Systems, Minneapolis, MN, United States), according to manufacturer’s instructions.
Alqahtani S., Kobos L.M., Xia L., Ferreira C., Franco J., Du X, & Shannahan J.H. (2020). Exacerbation of Nanoparticle-Induced Acute Pulmonary Inflammation in a Mouse Model of Metabolic Syndrome. Frontiers in Immunology, 11, 818.
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