Tnf alpha mouse elisa kit

Manufactured by Thermo Fisher Scientific

Sourced in United States

The TNF alpha Mouse ELISA Kit is a quantitative sandwich enzyme-linked immunosorbent assay (ELISA) designed for the measurement of mouse tumor necrosis factor alpha (TNF-α) in cell culture supernatants, serum, and plasma samples.

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18 protocols using tnf alpha mouse elisa kit

Isolation and Analysis of Mouse Peritoneal Macrophages

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The Mouse Peritoneal Macrophage Isolation Kit was purchased from Miltenyi Biotec (Bergisch Gladbach, Germany). An Enzyme-Linked Immunosorbent Assay (ELISA) Kit for evaluating PGE₂ was purchased from Abcam (Cambridge, Cambs, UK). The Mouse TNF-alpha ELISA Kit was purchased from eBioscience (San Diego, CA, USA). The Nuclear and Cytoplasmic Protein Extraction Kit was purchased from TransGen Biotech (Beijing, China).

Liu Y., Fang S., Li X., Feng J., Du J., Guo L., Su Y., Zhou J., Ding G., Bai Y., Wang S., Wang H, & Liu Y. (2017). Aspirin inhibits LPS-induced macrophage activation via the NF-κB pathway. Scientific Reports, 7, 11549.

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Electroacupuncture Therapy Mechanisms Explored

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Electroacupuncture Treatment Device (SDZ-II, Huatuo Brand) was from Suzhou Medical Appliance Factory, China. The size of the needles was 0.35 mm (Diameter) × 40 mm (Length). Antibodies used for immunostaining in this study were purchased from Abcam (MA, USA) including anti-CD34 (Mouse monoclonal, catalog no. ab8536), anti-VEGFA (Mouse monoclonal, catalog no. ab1316), anti-VEGF receptor-2 (Rabbit polyclonal, catalog no. ab2349), anti-angiopoietin-1(Rabbit polyclonal, catalog no. ab102015), anti-DLL4 (Rabbit polyclonal, catalog no. ab217860), anti-MMP-2 (Mouse monoclonal, catalog no. ab86607), anti-TIMP-2 (Mouse monoclonal, catalog no. ab1828), anti-integrin-β (Mouse monoclonal, catalog no. ab24693), anti-p75 NGF receptor (Rabbit monoclonal, catalog no. ab52987), anti-Sema3A (Rabbit polyclonal, catalog no. ab23393) and anti-Neuropilin 1 (Rabbit monoclonal, catalog no. ab81321). Mouse TNF-alpha ELISA kit was obtained from eBioscience (Thermo Fisher, MA, USA).

Jiang X., Tian Y., Xu L., Zhang Q., Wan Y., Qi X., Li B., Guo J., Sun W., Luo A., Huang J, & Gu X. (2019). Inhibition of Triple-Negative Breast Cancer Tumor Growth by Electroacupuncture with Encircled Needling and Its Mechanisms in a Mice Xenograft Model. International Journal of Medical Sciences, 16(12), 1642-1651.

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Quantifying TNFα Secretion in BV-2 Cells

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TNFα secretion was measured using a mouse TNF alpha ELISA kit (eBioscience) according to the manufacturer's instructions. Capture antibody was added in each well of a 96-well ELISA plate and incubated overnight at 4 °C. The plate was washed with buffer and then incubated with the supernatant (100 µl) from BV-2 cells exposed to different treatments for 2 hours at room temperature followed by 5 washes. Biotin-conjugated antibody was applied to each well and incubated for 30 min at room temperature followed by 5 washes. Diluted avidin-HRP was added, and the plate was incubated at room temperature for 30 min. After washing the plate, substrate solution was added to each well and the color was developed in the dark for 5 minutes at room temperature. The stop solution was added to each well, and the plate was read at 450 nm using a microplate reader. The concentration of TNFα was calculated using a mouse TNF alpha standard.

Markoutsa E, & Xu P. (2017). Redox potential sensitive N-acetyl cysteine-prodrug nanoparticles inhibit the activation of microglia and improve neuronal survival. Molecular pharmaceutics, 14(5), 1591-1600.

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FMP Modulates Inflammatory Cytokines

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RAW 264.7 cells were plated at a density of 1.5 × 10⁵ cells/well in 24-well plates and incubated for 24 h. Cells were treated with varying concentrations of FMP (12.5, 25, 50, and 100 µM) (then with or without LPS (1 µg/mL)) for 24 h. The concentrations of proinflammatory cytokines (TNF-α, IL-1β, and IL-6) in culture supernatant were determined using ELISA kits (Mouse TNF alpha ELISA Kit, Invitrogen, Carlsbad, CA, USA; Mouse IL-6 ELISA Kit, BD, Franklin Lakes, NJ, USA; Mouse IL-1β/IL-1F2, R&D Systems, Minneapolis, MN, USA).

Kim M.S., Park J.S., Chung Y.C., Jang S., Hyun C.G, & Kim S.Y. (2019). Anti-Inflammatory Effects of Formononetin 7-O-phosphate, a Novel Biorenovation Product, on LPS-Stimulated RAW 264.7 Macrophage Cells. Molecules, 24(21), 3910.

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Cytokine Response to 7AC and LPS

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RAW 264.7 cells were plated at a density of 1.0 × 10⁵ cells/well in 24-well plates and incubated 24 h. Cells were treated with varying concentrations of 7AC (50, 100 and 200 µM) with or without LPS (1 µg/mL) for 24 h. The concentrations of pro-inflammatory cytokines (TNF-α, IL-1β and IL-6) in culture supernatant were determined using respective ELISA kits (Mouse TNF alpha ELISA Kit, Invitrogen, Carlsbad, CA, USA; Mouse IL-6 ELISA Kit, San Diego, BD, USA; Mouse IL-1β/IL-1 F2, R&D Systems, Minneapolis, MN, USA).

Park T., Park J.S., Sim J.H, & Kim S.Y. (2020). 7-Acetoxycoumarin Inhibits LPS-Induced Inflammatory Cytokine Synthesis by IκBα Degradation and MAPK Activation in Macrophage Cells. Molecules, 25(14), 3124.

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Retinal Cytokine and Inflammation ELISA

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For the detection of inflammatory cytokine TNF‐a, IL‐1β, and IL‐6 in the retina, mouse TNF alpha Elisa Kit (Invitrogen), mouse IL‐1 beta Elisa Kit (Invitrogen), and mouse IL‐6 Elisa Kit (Invitrogen) were used based on the manufacturer's instructions. Optical density was measured at 450 nm on microplate reader (Azure Biosystems). The level of cAMP in the retina after plasma or recombinant TPM1 (rTPM1) protein treatment was evaluated with Cyclic AMP Competitive ELISA Kit (Invitrogen) or Camp ELISA Kit (Mouse) (Aviva Systems Biology). The expression levels of Cox‐1 and Cox‐2 in the retina were measured after rTPM1 protein treatment using mouse COX1 ELISA Kit (MyBioSource) and mouse COX2 ELISA Kit (Abcam), respectively.

Li R., Liang Y, & Lin B. (2022). Accumulation of systematic TPM1 mediates inflammation and neuronal remodeling by phosphorylating PKA and regulating the FABP5/NF‐κB signaling pathway in the retina of aged mice. Aging Cell, 21(3), e13566.

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Quantifying TNF-α and CXCL1 in Mice

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The levels of TNF‐α and CXCL1 proteins were measured using the TNF alpha Mouse ELISA Kit (Invitrogen) and Mouse CXCL1/KC Quantikine ELISA Kit (R&D System; Minneapolis, MN, USA), respectively. Assays were performed according to manufacturers’ instructions.

Ji J., Ding K., Cheng B., Zhang X., Luo T., Huang B., Yu H., Chen Y., Xu X., Lin H., Zhou J., Wang T., Jin M., Liu A., Yan D., Liu F., Wang C., Chen J., Yan F., Wang L., Zhang J., Yan S., Wang J., Li X, & Chen G. (2024). Radiotherapy‐Induced Astrocyte Senescence Promotes an Immunosuppressive Microenvironment in Glioblastoma to Facilitate Tumor Regrowth. Advanced Science, 11(15), 2304609.

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Cytokine Profiling in Mice

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The Lipocalin-2/NGAL DuoSet ELISA, mouse (R&D Systems), IL-18 ELISA Kit (Sino Biological) and TNF alpha Mouse ELISA Kit (invitrogen) were performed according to manufacturer’s instruction.

Fischer F., Romero R., Hellhund A., Linne U., Bertrams W., Pinkenburg O., Eldin H.S., Binder K., Jacob R., Walker A., Stecher B., Basic M., Luu M., Mahdavi R., Heintz-Buschart A., Visekruna A, & Steinhoff U. (2020). Dietary cellulose induces anti-inflammatory immunity and transcriptional programs via maturation of the intestinal microbiota. Gut Microbes, 12(1), 1829962.

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Analyzing Cytokine Secretion in Sensitized Cells

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RBL-2H3 cells (8 × 10⁵ cells/well) were sensitized overnight with 20 ng/mL DNP-IgE. BMMCs (1.0 × 10⁶ cells/well) were similarly sensitized with 100 ng/mL of DNP-specific IgE for 4 h. Subsequently, the cells were thoroughly washed with fresh culture medium, and then treated with nifuratel or PP2 for 30 min. The cells were then stimulated with 25 ng/mL (for RBL-2H3 cells) or 50 ng/mL (for BMMCs) DNP-HSA for 4 h at 37°C. The concentrations of TNF-α and IL-4 in the resulting culture medium were measured using Rat OptEIA ELISA kits from BD Biosciences (San Jose, CA, USA) or TNF alpha Mouse ELISA Kit from Invitrogen (Waltham, MA, USA) according to the manufacturer’s protocol.

Lee J.E., Choi M.Y., Min K.Y., Jo M.G., Kim Y.M., Kim H.S, & Choi W.S. (2023). Drug repositioning of anti-microbial agent nifuratel to treat mast cell-mediated allergic responses. International Journal of Immunopathology and Pharmacology, 37, 03946320231202349.

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Quantification of Inflammatory Cytokines

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TNFα in mouse serum was quantified with a TNF alpha Mouse ELISA Kit (BMS607-3, Thermo Fisher Scientific). IL-1β in mouse serum was quantified with a Mouse IL-1β ELISA Kit (RAB0274-1KT, Sigma-Aldrich). IL-6 in mouse serum was quantified with an IL-6 Mouse ELISA Kit (BMS603-2, Thermo Fisher Scientific). All assays were performed according to the instructions provided by the manufacturer.

Franko N., da Silva Santinha A.J., Xue S., Zhao H., Charpin-El Hamri G., Platt R.J., Teixeira A.P, & Fussenegger M. (2024). Integrated compact regulators of protein activity enable control of signaling pathways and genome-editing in vivo. Cell Discovery, 10, 9.

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