Tnf alpha human elisa kit

Enzyme‐linked immunosorbent assay (ELISA) was adopted to assess the levels of the inflammatory cytokines and the cell adhesion molecules in serum of CHD patients. Concretely, the inflammatory cytokines included tumor necrosis factor alpha (TNF‐α), interleukin (IL)‐1β, IL‐6, IL‐10, and IL‐17A; the cell adhesion molecules included VCAM‐1 and ICAM‐1. Commercial ELISA kits (Invitrogen, Carlsbad, California, USA) were applied for the assay, including TNF alpha Human ELISA Kit (Catalog# KAC1751), IL‐1 beta Human ELISA Kit (Catalog# KAC1211), IL‐6 Human ELISA Kit (Catalog# KAC1261), IL‐10 Human ELISA Kit (Catalog# KAC1321), IL‐17A Human ELISA Kit (Catalog# KAC1591), VCAM‐1 Human ELISA Kit (Catalog# KHT0601), and ICAM‐1 Human ELISA Kit (Catalog# BMS241). Assay procedures were implemented in strict accordance with experiment protocol provided by the manufacturer. The absorbance at 450 nm was read after the addition of stop solution immediately. A standard curve was performed with each assay, and the concentration of tested samples was read from the standard curve.

CD100 level in the plasma and supernatants was measured by human sCD100 ELISA kit (CUSABIO, Wuhan, China). MMP-2 level in the plasma was measured by MMP-2 Human ELISA kit (Invitrogen, ThermoFisher, Carlsbad, CA, USA). IFN-γ and TNF-α level in the supernatants was measured by IFN gamma Human ELISA kit (Invitrogen, ThermoFisher) and TNF alpha Human ELISA kit (Invitrogen, ThermoFisher), respectively.

Serum samples were assayed for the presence of biomarkers of inflammation (TNFα, CRP, IL-6), coagulation (Thrombin, D-dimer), oxidative stress (carbonyls and nitrites/nitrates), and Syndecan-1 as a marker of endothelial damage. All molecules were measured using commercially available kits following manufacturer instructions. CRP, D-Dimers, and IL-6 were measured by standardized central laboratory methodology. TNF-α was determined using the TNF alpha Human ELISA Kit (In Vitrogen catalog # KHC3011, Waltham, MA, USA). The measurement of thrombin was performed using the Human Thrombin ELISA Kit (Abcam, catalog # ab270210 Cambridge, U.K.). Protein carbonylation was evaluated using the Protein Carbonyl Fluorometric Assay Kit (Cayman Chemical, catalog # 701530 Ann Arbor, MI, USA). Total nitrites/nitrates concentration was determined using the Nitrate/Nitrite Fluorometric Assay Kit (Cayman Chemical, Catalog # 780051, Ann Arbor, MI, USA). Syndecan 1 (SDC1) was measured using Syndecan 1 Human ELISA Kit (Thermo Fisher Scientific, Catalog # EHSDC1, Waltham, MA, USA), Each sample was evaluated by duplicate, and the data were expressed as mean ± SD. Hemolyzed serums were discarded due to interference with the detection method.

Plasma levels of IL-6, IL-10, NF-κB, TNFα, and IFN-γ were measured using commercially available kits (IL-6 Human ELISA Kit, IL-10 Human ELISA Kit, NF kappaBp65 (Total) Human ELISA Kit, TNF alpha Human ELISA Kit, IFN gamma Human ELISA Kit, Invitrogen by Thermo (Fisher Scientific, Waltham, MA, USA) and the VersaMax Microplate Reader (Molecular Devices, San Jose, CA, USA).