Determiner l tg 2 kit

The blood glucose levels were determined using a Glutest Neo Super, Glutest Mint (Sanwa Chemical Co., Japan) or Glucose Assay Kit (BioVision, CA). Serum insulin levels and the triglyceride content in the liver were determined using an insulin kit (Morinaga, Japan) and the Determiner-L TG II kit (Wako Pure Chemical Industries, Japan), respectively. The levels of plasma alanine aminotransferase, free fatty acid, total cholesterol, and triglyceride were assayed using enzymatic methods (Wako Pure Chemical Industries). Serum DPP-4 activity was measured using a DPP4 Activity Assay Kit (Bio Vision, CA). Active GLP-1 was assayed using a Glucagon-Like Peptide-1 (Active) ELISA Kit (Millipore, MA). An insulin tolerance test (ITT) was performed by intraperitoneally injecting mice with human insulin (1.5 mU/g body weight). An oral glucose tolerance test (OGTT) was performed by withholding all food from the mice for more than 18 h and then orally loading the mice with glucose (1.5 mg/g body weight).

The blood glucose levels, serum insulin levels, triglyceride (TG) content in the liver, and glycogen content in the liver were determined using a Glutest Neo Super (Sanwa Chemical Co., Tokyo, Japan), an insulin kit (Morinaga, Tokyo, Japan), a leptin kit (Morinaga) and the Determiner-L TG II kit and the Determiner-GL-E kit (Wako Pure Chemical Industries, Osaka, Japan), respectively. The plasma alanine aminotransferase, free fatty acids, total cholesterol, LDL-cholesterol, and triglyceride levels were assayed using enzymatic methods (Wako Pure Chemical Industries). The plasma 3-hydroxybutyrate level was determined using the β Hydroxybutyrate Assay kit (Abcam, Cambridge, England). The blood glucose levels were checked just before and 4 hours after the administration of OSI-906 or the vehicle until day 7 or on days 8, 10, 14, 21, and 28. Blood samples were taken from the inferior vena cava immediately after sacrifice.