Mouse tnf alpha elisa ready set go

Manufactured by Thermo Fisher Scientific

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The Mouse TNF alpha ELISA Ready-SET-Go is a sandwich enzyme-linked immunosorbent assay (ELISA) kit designed to quantitatively measure mouse tumor necrosis factor alpha (TNF-alpha) levels in biological samples.

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11 protocols using mouse tnf alpha elisa ready set go

Quantification of Cytokines in Cell Culture

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Philipp D., Suhr L., Wahlers T., Choi Y.H, & Paunel-Görgülü A. (2018). Preconditioning of bone marrow-derived mesenchymal stem cells highly strengthens their potential to promote IL-6-dependent M2b polarization. Stem Cell Research & Therapy, 9, 286.

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Cytokine Quantification in Cell Culture and Plasma

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For cytokine quantification in culture supernatants, the following ELISA kits have been used: Human/mouse TGF beta 1 2nd Generation ELISA Ready-SET-Go (eBioscience), Mouse IL-6 DuoSet ELISA (R&D Systems), Mouse IL-10 DuoSet ELISA (R&D Systems) and Mouse TNF alpha ELISA Ready-SET-Go (eBioscience) following the manufacturers' instructions. Cardiac troponin (cTnT) was quantified using Mouse cTnT/TNNT2 ELISA Kit (Immunoway).
A mouse multiplex Immunoassay (ProcartaPlex 4 Plex, Thermo Fisher) and Luminex 200 System (Thermo Fisher) were used to quantify cytokines in plasma samples.

Eghbalzadeh K., Georgi L., Louis T., Zhao H., Keser U., Weber C., Mollenhauer M., Conforti A., Wahlers T, & Paunel-Görgülü A. (2019). Compromised Anti-inflammatory Action of Neutrophil Extracellular Traps in PAD4-Deficient Mice Contributes to Aggravated Acute Inflammation After Myocardial Infarction. Frontiers in Immunology, 10, 2313.

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Quantifying Inflammatory Cytokines in Cells

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IL-6, TNF and IL-1beta levels in cell-culture supernatants were determined using a Mouse IL-6 ELISA Ready-SET-Go, Mouse TNF alpha ELISA Ready-SET-Go, and Mouse IL-1b ELISA Ready-SET-Go kits (eBioscience, San Diego, CA, USA) according to the manufacturer's instructions.

Nosenko M.A., Moysenovich A.M., Zvartsev R.V., Arkhipova A.Y., Zhdanova A.S., Agapov I.I., Vasilieva T.V., Bogush V.G., Debabov V.G., Nedospasov S.A., Moisenovich M.M, & Drutskaya M.S. (2018). Novel Biodegradable Polymeric Microparticles Facilitate Scarless Wound Healing by Promoting Re-epithelialization and Inhibiting Fibrosis. Frontiers in Immunology, 9, 2851.

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Quantification of Cytokines in Cell Culture

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IL-6 and TNF levels in cell-culture supernatants were determined using a Mouse IL-6 ELISA Ready-SET-Go and Mouse TNF alpha ELISA Ready-SET-Go kits (eBioscience, San Diego, CA, USA) according to the manufacturer's instructions.

Korneev K.V., Kondakova A.N., Sviriaeva E.N., Mitkin N.A., Palmigiano A., Kruglov A.A., Telegin G.B., Drutskaya M.S., Sturiale L., Garozzo D., Nedospasov S.A., Knirel Y.A, & Kuprash D.V. (2018). Hypoacylated LPS from Foodborne Pathogen Campylobacter jejuni Induces Moderate TLR4-Mediated Inflammatory Response in Murine Macrophages. Frontiers in Cellular and Infection Microbiology, 8, 58.

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Quantifying Cytokine Levels in Mice

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The cytokine level of serum of WT and KI mice were measured using commercial ELISA kits: mouse TNF alpha ELISA Ready-SET-Go (eBioscience, Cat# 88-7324-77) and mouse IL-6 ELISA Ready-SET-Go (eBioscience, Cat# 88-7064-88) following the manufacturers’ instructions.

Zheng G., Jiang C., Li Y., Yang D., Ma Y., Zhang B., Li X., Zhang P., Hu X., Zhao X., Du J, & Lin X. (2018). TMEM43-S358L mutation enhances NF-κB-TGFβ signal cascade in arrhythmogenic right ventricular dysplasia/cardiomyopathy. Protein & Cell, 10(2), 104-119.

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Mouse TNF-alpha ELISA Protocol

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TNF-α levels were determined in duplicate in 100 µL samples, using a commercially available ELISA kit (Mouse TNF alpha ELISA Ready-SET-Go!, eBioscience™, San Diego, CA, USA). The absorbance was measured spectrophotometrically at 450 nm (TECAN Sunrise™ powered by Magellan™ data analysis software, Mannedorf, Switzerland). The assays sensitivity was 8 pg/mL.

Giorgio C., Allodi M., Palese S., Grandi A., Tognolini M., Castelli R., Lodola A., Flammini L., Cantoni A.M., Barocelli E, & Bertoni S. (2021). UniPR1331: Small Eph/Ephrin Antagonist Beneficial in Intestinal Inflammation by Interfering with Type-B Signaling. Pharmaceuticals, 14(6), 502.

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Quantification of TNF-α in Mouse Cell Cultures

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At the end of the incubation period with the test compounds and in the absence or presence of ionomycin and PMA, the culture media were frozen at −80°C until cytokine measurement. TNF-α levels were determined in duplicate in 100 µL samples, using a commercially available ELISA kit (Mouse TNF alpha ELISA Ready-SET-Go!, eBioscience^™, San Diego, CA). The absorbance was measured spectrophotometrically at 450 nm (TECAN Sunrise^™ powered by Magellan^™ data analysis software, Mannedorf, Switzerland). The assays sensitivity was 8 pg/mL.

Grandi A., Zini I., Palese S., Giorgio C., Tognolini M., Marchesani F., Bruno S., Flammini L., Cantoni A.M., Castelli R., Lodola A., Fusari A., Barocelli E, & Bertoni S. (2019). Targeting the Eph/Ephrin System as Anti-Inflammatory Strategy in IBD. Frontiers in Pharmacology, 10, 691.

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Stimulating Type I Interferon Response

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Recombinant human IFNβ (R&D, cat no. 8499-IF-010/CF), diABZI (Invivogen, cat no. 2138299-34-8), 2’3’-cGAMP (Invivogen, cat no. tlrl-nacga23-02), DMXAA (Invivogen, cat no. tlrl-dmx), H151 (Invivogen, cat no. inh-H151), RpRpSSCDA (Biolog, cat no. C118), THP1-Dual^™ KO-STING Cells (Invivogen, cat no. thpd-kostg). All other chemical inhibitors are from Selleckchem. Mouse TNF alpha ELISA Ready Set Go. (eBioscience, cat no. 88–7324). Mouse IFN-Beta ELISA Kit (PBI, cat no. 42400).

Aybar-Torres A., Saldarriaga L.A., Pham A.T., Emtiazjoo A.M., Sharma A.K., Bryant A.J, & Jin L. (2024). The common TMEM173 HAQ, AQ alleles rescue CD4 T cellpenia, restore T-regs, and prevent SAVI (N153S) inflammatory disease in mice. bioRxiv.

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Murine Cytokine Quantification ELISA

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Murine IL-6 and TNF levels in cell-culture supernatants were measured using a Mouse IL-6 ELISA Ready-SET-Go! and Mouse TNFalpha ELISA Ready-SET-Go! kits (eBioscience, San Diego, CA, USA) according to the manufacturer’s protocol.

Korneev K.V., Arbatsky N.P., Molinaro A., Palmigiano A., Shaikhutdinova R.Z., Shneider M.M., Pier G.B., Kondakova A.N., Sviriaeva E.N., Sturiale L., Garozzo D., Kruglov A.A., Nedospasov S.A., Drutskaya M.S., Knirel Y.A, & Kuprash D.V. (2015). Structural Relationship of the Lipid A Acyl Groups to Activation of Murine Toll-Like Receptor 4 by Lipopolysaccharides from Pathogenic Strains of Burkholderia mallei, Acinetobacter baumannii, and Pseudomonas aeruginosa. Frontiers in Immunology, 6, 595.

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Measuring Cytokine Levels in pDCs

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For cytokine measurement in culture supernatants, spleen or BM cells from individual mice in the same group were pooled prior to FACS purification. FACS-purified pDCs (3 × 10⁴ cells/well) were incubated in 120ul RPMI-1640 supplemented with 10% (vol/vol) fetal bovine serum, L-glutamine, penicillin-streptomycin, and HEPES pH 7.2) supplemented with 50μM β-mercaptoethanol in the presence or absence of CpG-B 1668 (Integrated DNA Technologies) at 0.1μM (Flt3L culture-derived pDC) or 1μM (splenic and BM pDC) or CpG-A 1585 (InvivoGen) at 0.5uM for 6 or 15 hours at 37°C. Supernatants were collected and stored at −80°C until further analysis. IFN-I bioactivity was measured with reference to a recombinant mouse IFNβ standard (Sigma-Aldrich) using ISRE-L929 reporter cells (L-929 cell line transfected with an interferon-sensitive luciferase) (Jiang et al., 2005 (link)). IFNα and IFNβ were measured by Lumikine mIFNα and Lumikine Xpress mIFNβ ELISA (InvivoGen), respectively, following manufacturer instructions. TNFα was measured by Mouse TNF alpha ELISA Ready-SET-Go!® (eBioscience) following manufacturer instructions. Graphs depicting cytokine measurements in this paper represent individual wells of stimulation.

Macal M., Jo Y., Dallari S., Chang A.Y., Dai J., Swaminathan S., Wehrens E., Fitzgerald-Bocarsly P, & Zúñiga E.I. (2018). Self-renewal and Toll-like receptor Signaling Sustain Exhausted Plasmacytoid Dendritic Cells During Chronic Viral Infection. Immunity, 48(4), 730-744.e5.

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