Icam1 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.

Sub-confluent HGFs were pre-treated with 6-shogaol (5 or 10 μM) for 1 h and cultured with AGEs (500 μg/mL) or BSA for 48 h. NAC (1 mM) was also added to HGF cultures with AGEs. The conditioned medium was collected and mixed with protease inhibitor cocktail, and cell lysates were extracted in lysis buffer including 10 mmol/l Tris-HCl, pH 7.4, 50 mmol/l NaCl, 5 mmol/l EDTA, 1 mmol/l sodium orthovanadate, 1% NP-40 and protease inhibitor cocktail. IL-6 in the conditioned medium was measured using a Human IL-6 ELISA kit (R&D Systems, Minneapolis, MN, USA) in accordance with the manufacturer’s instructions. ICAM-1 in the cell lysate was determined using a ICAM1 Human ELISA kit (Invitrogen, Carlsbad, CA, USA), and the amount of ICAM-1was normalized to total cell protein amount.