Microplate scanning spectrometer

Six week old mice were sacrificed and plasma leptin and cholecystokinin were measured using the Quantikine ELISA mouse Leptin Immunoassay kit (R&D Systems) and the Quantikine ELISA mouse Cholecystokinin ELISA Immunoassay kit (R&D systems), respectively, according to the manufacturer’s protocol. A microplate scanning spectrometer (BioTek) was used to measure the optical density for leptin and CCK (450 nm wavelength correction: 540 nm–570 nm).
Purified leptin (R&D Systems) and CCK (Bachem) were administered to six week old mice intraperitoneally at doses of 5 mg/kg and 10 ug/kg, respectively.

Plasma leptin, adiponectin, cholesterol, and insulin were measured in both CD and HFD mice using the following commercially available kits; the Quantikine^® ELISA mouse Leptin Immunoassay kit (Catalog No: MOB00; R & D Systems, Minneapolis, MN, USA), the Quantikine^® ELISA mouse Adiponectin/Acrp30 Immunoassay kit (Catalog No: MRP300, R & D systems, Minneapolis, MN, USA), cholesterol assay kit (Catalog No: ab65390; Abcam, Cambridge, MA, USA) and mouse insulin (INS) ELISA kit (Catalog No: E-EL-M1382; Elabscience^®, Houston, TX, USA), respectively, following the manufacturer’s protocol. A microplate scanning spectrometer (BioTek instruments Inc., Highland, CA, USA) was used to measure the optical density for the leptin (450 nm, wavelength correction: 540 nm–570 nm), adiponectin (450 nm, wavelength correction: 540 nm–570 nm) and insulin (450 nm) assays. A fluorometric detection method was used to measure total cholesterol, and optical density (570 nm) was measured on a luminescence spectrometer.