Cytokines were quantified using the following ELISA kits: the IL-1β Quantikine ELISA kit (R&D Systems, Minneapolis, MN), IL-6 Quantikine ELISA kit (R&D Systems, Minneapolis, MN), and TNF-α Quantikine HS ELISA Kit (R&D Systems, Minneapolis, MN). The skin tissue was excised and homogenized in PBS. The number of viable bacterial cells in the tissue was determined by counting the number of colonies formed on mannitol salt medium (Nissui Pharmaceutical, Tokyo, Japan) supplemented with 10 µg/mL ampicillin. Excised tissues were homogenized in lysis buffer 2 (R&D Systems, Minneapolis, MN). After preparation according to the manufacturers’ instructions, lysates were stored at − 80 °C until the quantification of IL-1β. Blood samples were obtained via cardiac puncture under inhalation anaesthesia. After incubation for two hours at room temperature, serum was prepared and stored at − 80 °C until the quantification of IL-6 and TNF-α.
Tnf α quantikine hs elisa kit
Manufactured by R&D Systems
The TNF-α Quantikine HS ELISA Kit is a quantitative sandwich enzyme immunoassay designed for the measurement of human tumor necrosis factor alpha (TNF-α) concentrations in cell culture supernates, serum, and plasma. The kit utilizes a microplate pre-coated with a monoclonal antibody specific for TNF-α.
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2 protocols using tnf α quantikine hs elisa kit
1
Quantifying Cytokine Levels in Skin Tissue
2
Measurement of Biomarkers in Cardiovascular Disease
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Malondialdehyde was determined spectrophotometrically with a commercial kit (Oxford Biomedical Research, Rochester Hills, MI; colorimetric assay for lipid peroxidation; measurement range, 1-20 nmol/L). 15 (link) IL-6, IL-10, IL-12, and TNF-α were measured using high-sensitivity immunoassays (human IL-6, IL-10, IL-12, TNF-α Quantikine HS ELISA Kit; R&D Systems, Inc, Minneapolis, MN). These assays detect values as low as 0.11 pg/mL for IL-6, 0.17 pg/ mL for IL-10, 0.5 pg/mL for IL-12, and 0.191 pg/mL for TNF-α, respectively. Fetuin-a (Human Fetuin-A ELISA Kit; BioVendor -Laboratory Medicine, Brno, Czech Republic) and IL-17A (Human IL-17A High Sensitivity ELISA; eBioscence, Vienna, Austria) were also measured by ELISA with sensitivity of 0.104 ng/mL and 0.01 pg/mL, respectively. NT-proBNP was measured using the Elecsys NT-proBNP-Cobas (Roche Diagnostics, Rotkreuz, Switzerland) system with sensitivity of 5 ng/L. The intra-assay coefficient of variation was <5% for all tests.
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