Mice plasma samples were analysed for the concentrations of insulin (Mouse Insulin ELISA, ALPCO, USA, 80-INSMS-E01, E10), TNF-α (Mouse TNF-α ELISA, BioLegend, USA, 430901), IL-6 (Mouse IL-6 ELISA, BioLegend, USA, 431301) and IL-10 (Mouse IL-10 ELISA, BioLegend, USA, 431411) as per manufacturer instructions.
Mouse tnf α elisa
Manufactured by BioLegend
Sourced in United States
The Mouse TNF-α ELISA is a quantitative sandwich enzyme-linked immunosorbent assay designed for the measurement of mouse tumor necrosis factor-alpha (TNF-α) levels in cell culture supernatants, serum, and plasma.
Automatically generated - may contain errors
Lab products found in correlation
Mouse il 10 elisa, by BioLegend (2 mentions)
Microplate reader, by Thermo Fisher Scientific (2 mentions)
Mouse insulin elisa, by ALPCO (1 mentions)
Il 6 mouse elisa, by BioLegend (1 mentions)
Il 15, by BioLegend (1 mentions)
Il 18, by Elabscience (1 mentions)
Il 12, by BioLegend (1 mentions)
Rat tnf α duoset elisa, by R&D Systems (1 mentions)
Il 6 elisa kit, by BioLegend (1 mentions)
3 3 5 5 tetramethylbenzidine (tmb), by Merck Group (1 mentions)
4 protocols using mouse tnf α elisa
1
Cytokine Profiling in Mouse Plasma
2
Cytokine Measurement in Tumor and Macrophage Samples
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The ELISA assays used 50 μl of culture supernatant after centrifugation at 16,000 g at 4°C for 10 min. The mouse cytokines IL-12 (Biolegend 431704), Il-15 (Biolegend 435104), and IL-18 (Elabscience) were used according to the manufacturers’ instructions. Mouse IL-10 ELISA (Biolegend, 431414), Mouse TNFα ELISA (Biolegend, 430901), and Mouse IL-12p70 ELISA (Biolegend, 433604) were used for intratumor cytokine analysis. For the THP-1 supernatant experiment, Human IL-10 ELISA (Biolegend, 430604) and human IL-6 ELISA (Biolegend, 430504) were used for macrophage polarization analysis. The chemiluminescent signals were measured at 450 nm.
3
BV2 Microglia Inflammation Assay
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BV2 murine microglia cells (4×105) were seeded in 35 mm dishes. Next day, the cells were treated with GSK2578215A (hereinafter GSK, 1 µM, Torcris Bioscience) for 30 min followed by LPS (1 µg/ml) treatment for 6 h. Then, the cells was fixed with 4% paraformaldehyde (PFA, Wako) for immunofluorescence analysis or harvested for Western blot analysis. Culture media derived from these BV2 cells and rat primary microglia administrated with the same treatment as BV2 cells, and brain lysates from G2019S transgenic mouse or littermate were used to perform mouse TNFα ELISA (Biolegend) or rat TNFα DuoSet ELISA (R&D systems) according to the sample species and the manufacturer's instruction.
4
Quantification of Inflammatory Cytokines
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The amount of TNF-α and IL-6 was quantified in culture supernatants from BMDMs treated as indicated using mouse TNF-α ELISA and IL-6 ELISA kits (BioLegend, CA, USA) following the manufacturer’s instructions. Streptavidin HRP was used to detect bound antibodies, and TMB (Sigma Aldrich) was used as a substrate. The reaction was stopped with 1 M H2SO4. Absorbance at 450 and 620 nm was measured on a microplate reader (Thermo Fisher Scientific).
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