Ffa quantitation kit

Body composition was examined using an EchoMRI system. For serum FFA level determination, tail-vein blood was collected at designated times and FFA levels were measured using an FFA Quantitation Kit (Sigma-Aldrich). For GTT, mice were fasted overnight and injected intraperitoneally with glucose (1.5 g/kg body weight), then tail-vein blood was collected at designated times, and blood glucose levels were measured using a Nova Max Glucometer. For ITT, mice were fasted for 6 h in the morning and injected intraperitoneally with insulin (1 U/kg body weight for NC-fed mice; 1.5 U/kg body weight for HFD-fed mice), then blood glucose levels were measured as described above. For metabolic cage analysis, mice were acclimated in metabolic chambers for 3 days and then gas exchange, energy expenditure, food intake, and physical activity were recorded continuously for 4 days.

After intraperitoneal injection of heparin-sodium, mouse blood was collected by cardiac puncture under anesthesia as previously described [25 (link)]. Serum cholesterol, alanine aminotransferase and aspartate aminotransferase levels were measured using the Reflotron^® Plus System (Roche Diagnostics, Rotkreuz, Switzerland). Measurement of serum FFA was performed using the FFA quantitation kit (Sigma-Aldrich, St. Louis, USA).

Mouse serum was obtained after the 30 days of sulpiride treatment. PRL was evaluated by ELISA as previously reported [34 (link)]; insulin was measured using the Ultra Sensitivity Mouse Insulin ELISA Kit (90080, Crystal Chem, Elk Grove Village, IL); TG were measured using a colorimetric assay kit (10010303–96, Cayman Chem, Ann Arbor, MI); serum FFA was measured using the FFA Quantitation Kit (MAK044-1KT, Sigma-Aldrich, Burlington, MA); serum glycerol was measured using the Glycerol Assay Kit (MAK117-1KT, Sigma-Aldrich), and glucose levels were measured in fasting (4 h without food) and postprandial conditions (2 h without food) using glucose test strips (Accu-Check Active, 07124112, Roche, Mannheim, Germany). Glucagon levels were evaluated using a Mouse Glucagon ELISA kit (81518, Crystal Chem, Elk Grove Village, IL).