Il 10 mouse uncoated elisa kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The IL-10 Mouse Uncoated ELISA Kit is a laboratory product designed for the quantitative measurement of Interleukin-10 (IL-10) levels in mouse biological samples. It utilizes the Enzyme-Linked Immunosorbent Assay (ELISA) technique to detect and quantify the target analyte.

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11 protocols using il 10 mouse uncoated elisa kit

Splenic B cell IL-10 Induction

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Splenic CD19⁺ B cells were pre-sorted by the CD19 mAb-micro-beads (Miltenyi Biotec, Bergisch Gladbach, Germany). The sorted B cells (3 × 10⁶) were stimulated with or without LPS (10 μg/ml) and with or without entinostat for 48 h. The supernatants were harvested and measured by using the IL-10 Mouse Uncoated ELISA Kit (Invitrogen, Carlsbad, CA, USA) in accordance with the manufacturer’s instructions.

Min K.Y., Lee M.B., Hong S.H., Lee D., Jo M.G., Lee J.E., Choi M.Y., You J.S., Kim Y.M., Park Y.M., Kim H.S, & Choi W.S. (2021). Entinostat, a histone deacetylase inhibitor, increases the population of IL-10+ regulatory B cells to suppress contact hypersensitivity. BMB Reports, 54(10), 534-539.

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SARS-CoV-2 VLP-induced Cytokine Production

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TNF-α mouse uncoated enzyme-linked immunosorbent assay (ELISA) kit, IL-1β mouse uncoated ELISA kit, IL-10 mouse uncoated ELISA kit, IL-12 p70 mouse uncoated ELISA kit, interferon (IFN)-γ mouse uncoated ELISA kit, and IL-4 mouse uncoated ELISA kit were purchased from Invitrogen (ThermoFisher Scientific, USA). DCs (5 × 10⁵ cells/mL) were incubated with 5, 10, or 15 μg/mL SARS-CoV-2 VLPs for 24 h; 50 ng/mL lipopolysaccharide-treated DCs were used as the positive control. The culture supernatants were collected and evaluated for the levels of TNF-α, IL-1β, IL-10, IL-12p70, IFN-γ, and IL-4 using the respective ELISA kit according to the manufacturer’s instructions. The levels of cytokines released into culture medium were determined by measuring OD450 using a microplate reader.

Mi Y., Liang L., Xu K., Li Q., Wang W., Dang W., Deng J., Zhi Y., Li X, & Tan J. (2022). Severe acute respiratory syndrome coronavirus 2 virus-like particles induce dendritic cell maturation and modulate T cell immunity. Frontiers in Cellular and Infection Microbiology, 12, 986350.

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Inflammatory Cytokine Profiling of Adipose Tissue and Serum

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Serum samples were collected by cardiac puncture and collected in Microtainer tubes (365978, BD Bioscience). Samples were centrifuged for 90 s at 15,000 g and serum aliquots were snap‐frozen until further analysis. Global inflammatory cytokine analysis of supernatants of adipose tissue explant cultures and serum were performed using LEGENDPlex™ Mouse Inflammation Panel (740446, Biolegend). Supernatant IL‐10 levels were measured by IL‐10 Mouse Uncoated ELISA Kit (88‐7105‐86, Invitrogen). Adipose tissue‐derived cytokine levels were normalized to input tissue weight.

Richter F.C., Friedrich M., Kampschulte N., Piletic K., Alsaleh G., Zummach R., Hecker J., Pohin M., Ilott N., Guschina I., Wideman S.K., Johnson E., Borsa M., Hahn P., Morriseau C., Hammock B.D., Schipper H.S., Edwards C.M., Zechner R., Siegmund B., Weidinger C., Schebb N.H., Powrie F, & Simon A.K. (2023). Adipocyte autophagy limits gut inflammation by controlling oxylipin and IL‐10. The EMBO Journal, 42(6), e112202.

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C. costata Modulates Inflammatory Mediators

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The effect of C. costata fractions on the inflammatory (TNF-α and IL-6) and anti-inflammatory (IL-10) mediators was evaluated using the ELISA assay in RAW 264.7 cells treated with LPS. Cells were seeded in 12-well plates (5 × 10⁵ cells/well) and incubated for 18 h with C. costata fractions and LPS. The supernatant (1000 µL) was collected and the levels of TNF-α, IL-6 and IL-10 were quantified using the TNF Alpha Mouse uncoated ELISA Kit (# 88-7324-22, Invitrogen™, ThermoFisher Scientific, Waltham, MA, USA), IL-6 Ready-SET Go ELISA Kit (#88-7064-22, Invitrogen™, ThermoFisher Scientific) and IL-10 Mouse uncoated ELISA Kit (#88-7105-22, Invitrogen™, ThermoFisher Scientific), respectively, according to the manufacturer’s instructions. The absorbance was measured at 570 nm and 450 nm, to allow subtraction of the wavelength. TNF-α, IL-6 and IL-10 levels were expressed as a percentage of the control.

Susano P., Silva J., Alves C., Martins A., Gaspar H., Pinteus S., Mouga T., Goettert M.I., Petrovski Ž., Branco L.B, & Pedrosa R. (2021). Unravelling the Dermatological Potential of the Brown Seaweed Carpomitra costata. Marine Drugs, 19(3), 135.

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ELISA Quantification of IFN-γ and IL-10

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IFN-γ and IL-10 concentrations in cell culture supernatants were quantified by using the IFN gamma Mouse Uncoated ELISA Kit and the IL-10 Mouse Uncoated ELISA Kit (both Thermo Fisher Scientific), respectively, according to the manufacturer´s protocol. Samples were measured with the SUNRISE Absorbance Reader and analyzed utilizing the Magellan software (both Tecan Group, Männedorf, Switzerland).

Bauer V., Ahmetlić F., Hömberg N., Geishauser A., Röcken M, & Mocikat R. (2021). Immune checkpoint blockade impairs immunosuppressive mechanisms of regulatory T cells in B-cell lymphoma. Translational Oncology, 14(9), 101170.

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Cytokine Expression Quantification

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IL-12, IL-10, TGF-β, and IFN-γ were quantified using an IL-12 p70 Mouse Uncoated ELISA Kit (Thermo Fisher Scientific), IL-10 Mouse Uncoated ELISA Kit (Thermo Fisher Scientific), TGF beta-1 Human/Mouse Uncoated ELISA Kit (Thermo Fisher Scientific), and IFN gamma Mouse Uncoated ELISA Kit (Thermo Fisher Scientific), respectively, according to the manual of each kit.

Kajiyama S., Nagatake T., Ishikawa S., Hosomi K., Shimada Y., Matsui Y, & Kunisawa J. (2023). Lentinula Edodes Mycelia extract regulates the function of antigen-presenting cells to activate immune cells and prevent tumor-induced deterioration of immune function. BMC Complementary Medicine and Therapies, 23, 281.

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Cytokine-induced IL-10 Secretion

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Lin⁻ cells (1 × 10⁵ cells per well) sorted by using a lineage cell depletion kit (Miltenyi Biotec) were incubated in 96-well plates for 12 h in the presence of the indicated cytokines or IL-27 (1, 10 and 100 ng/ml; Biolegend). IL-10 levels were quantified using an IL-10 mouse uncoated ELISA kit (Thermo Fisher, Waltham, MA) according to the manufacturer’s instructions.

Min K.Y., Kim D.K., Jo M.G., Choi M.Y., Lee D., Park J.W., Park Y.J., Chung Y., Kim Y.M., Park Y.M., Kim H.S, & Choi W.S. (2024). IL-27-induced PD-L1highSca-1+ innate lymphoid cells suppress contact hypersensitivity in an IL-10-dependent manner. Experimental & Molecular Medicine, 56(3), 616-629.

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Macrophage Responses to SS-SF Materials

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The in vitro cell responses to the SS-SF materials were assessed using macrophages RAW264.7 (TIB-71, ATCC, Manassas, VA, USA). SS-SF film discs or fibroin film discs with 8 mms diameter sterilized by UV radiation for 30 min, conditioned with Dulbecco’s modified eagle medium (DMEM) for 6 h before cell seeding in 96 well plates. Aliquots (100 μL, 1×10⁴ cells) of RAW264.7 suspensions at ~90% confluence were seeded on the surfaces of the SS-SF films or fibroin films. Meanwhile, equal amounts of RAW264.7 cells were seeding directly into the wells of 96-well plates without films with or without lipopolysaccharide (LPS) at doses of 5 μg/mL as positive or negative controls, respectively. Cells were then cultivated in DMEM with 10% (w/v) FBS at 37°C and 5% CO₂. At days 1 and 7 post cell seeding, the cell medium supernatants were collected for quantitation of TNF-α and IL-2 release using an TNF alpha Mouse ELISA Kit (Thermo Fisher Scientific, USA) and IL-10 Mouse Uncoated ELISA Kit (Thermo Fisher Scientific, USA) according to the instructions of the manufacturer.

Wang F., Guo C., Li C., Zhao P., Xia Q, & Kaplan D.L. (2020). Protein composites from silkworm cocoons as versatile biomaterials. Acta biomaterialia, 121, 180-192.

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Quantification of IL-10 levels

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ALT and AST levels were quantified in the serum using an AutoAnalyzer (Prestige 24i, PZ Cormay S.A) as previously described [10] (link).
2.8. IL-10 quantification IL-10 levels were quantified on blood, spleen and liver homogenates by ELISA and the transcriptional levels of Il10 gene were assessed by real-Time quantitative PCR (qRT-PCR). Briefly, spleen and liver were homogenized and stored for IL-10 quantification using IL-10 mouse uncoated ELISA kit (ThermoScientific) or in TRIreagent (Sigma-Aldrich) for RNA extraction. Total RNA was extracted and the synthesis of cDNA was made with SensiFAST™ cDNA Synthesis Kit (Bioline). qRT-PCR reactions were run for each sample on a Bio-Rad CFX96 Real-Time System C1000 Thermal Cycler (Bio-rad) using SensiFAST SYBR Hi-ROX Kit (Bioline). Primer sequences were obtained from Alfagene (Portugal) and thoroughly tested. The results were normalized to the expression of the housekeeping gene ubiquitin. After amplification, cycle thresholdvalues (Ct-values) were calculated for all samples and gene expression changes were analyzed in the CFX Manager Software (Bio-Rad). The serum, splenic and liver IL-10 transcript and expression levels are shown on Supplementary figure 2A and B respectively.

Mesquita I., Ferreira C., Barbosa A.M., Ferreira C.M., Moreira D., Carvalho A., Cunha C., Rodrigues F., Dinis-Oliveira R.J., Estaquier J., Castro A.G., Torrado E, & Silvestre R. (2018). The impact of IL-10 dynamic modulation on host immune response against visceral leishmaniasis. Cytokine, 112.

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Cytokine Quantification in Microglia

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The levels of cytokines released from BV2 cells and primary microglia in culture were quantified using the following commercial enzyme-linked immunosorbent assay (ELISA) kits according to the manufacturer’s instructions: mouse TNF-α uncoated ELISA kit (88–7324-22, Thermo Fisher, USA), mouse IL-6 uncoated ELISA kit (88–7064-22, Thermo Fisher, USA), mouse IL-4 uncoated ELISA kit (88–7044-22, Thermo Fisher, USA), and mouse IL-10 uncoated ELISA kit (88–7105-22, Thermo Fisher, USA).

Feng L., Luo G., Li Y., Zhang C., Liu Y., Liu Y., Chen H., He D., Zhu Y, & Gan L. (2023). Curcumin-dependent phenotypic transformation of microglia mediates resistance to pseudorabies-induced encephalitis. Veterinary Research, 54, 25.

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