Ultrasensitive mouse insulin enzyme linked immunosorbent assay kit

In chow-fed mice, plasma albumin, bilirubin, blood urea nitrogen, creatinine, ALT, and AST levels were measured using the AU480 Clinical Chemistry Analyzer (Beckman Coulter, Providence, RI). In high-fat–fed mice, plasma ALT and AST activity was measured using the ALT Activity Assay Kit (700260; Cayman Chemical, Ann Arbor, MI) and the AST Activity Assay Kit (MAK055; Sigma-Aldrich, St. Louis, MO), respectively. Fasting blood glucose and plasma insulin were assessed using the Accu-Chek glucometer (Roche Diagnostics, Basel, Switzerland) and the Ultrasensitive Mouse Insulin Enzyme-Linked Immunosorbent Assay Kit (90080; Crystal Chem, Downers Grove, IL), respectively. TAG and collagen content were assessed in liver homogenate using the Triglyceride Quantification Colorimetric Kit (K622-100; BioVision, Mountain View, CA) and the Hydroxyproline Colorimetric Assay Kit (MAK008; Sigma-Aldrich), respectively. To characterize hepatic oxidative stress, 4-HNE and TBARS levels in liver homogenate were measured with the OxiSelect HNE Adduct Competitive Enzyme-Linked Immunosorbent Assay Kit (STA-838; Cell Biolabs, Inc, San Diego, CA) and the Lipid Peroxidation Assay Kit (MAK085; Sigma-Aldrich), respectively.

Glucose was measured from the tail-tip with a glucometer. The total RNA, including miRNAs, was extracted and purified using QIAzol lysis reagent and kits according to the manufacturer’s instructions (Qiagen, Hilden, Germany). Plasma insulin levels were measured using an ultrasensitive mouse insulin enzyme-linked immunosorbent assay kit (Crystal Chem, Downers Grove, Illinois).