Quantikine high sensitivity enzyme linked immunosorbent assay elisa kits

Serum glucose and triglyceride concentrations were measured by using enzymatic and colorimetric methods, respectively (AU5800, Beckman Coulter Inc., California, USA). Serum insulin was measured by using a chemiluminescence immunoassay (ADVIA Centaur, Siemens Healthcare Diagnostics, Hamburg, Germany). These analyses were carried out in a laboratory accredited by the College of American Pathologists. NEFA was measured at Mayo Medical Laboratories (Rochester, MN, USA), using an enzymatic colorimetric method (Cobas® 6000, Roche Diagnostics, Indianapolis, USA). Plasma IL-6 (cat no. HS600B) concentrations were measured using Quantikine® high-sensitivity enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems, Minneapolis, MN, USA). Plasma TNFα concentration was measured using an ultrasensitive ELISA kit (cat no. 45-TNFHUU-E01, Alpco Diagnostics, Salem, NH, USA). Intra- and inter-assay coefficient of variations for IL-6 and TNFα were largely within 10%.

Quantikine high sensitivity enzyme-linked immunosorbent assay (ELISA) kits (R&D Systems) were used to assess the expression of basic fibroblast growth factor (bFGF) and vascular endothelial growth factor (VEGF) in decellularized ECM according to the manufacturer's instructions. The absorbance of the wells at 450 nm was measured using a Multiskan MK3 microplate reader (Thermo Fisher Scientific, Waltham, Massachusetts). All samples were assayed in duplicate. A standard curve was constructed by plotting the mean absorbance of each standard against its concentration. The concentration of two growth factors was also normalized to the surface area of the culture dishes, and each growth factor amount was denoted as pg/cm².