Sensitive insulin ria kit

For each condition, 25 islets were transferred to medium containing 3.0 mM glucose for 3 hours to establish baseline insulin secretion. Next, 25 islets per condition were incubated for 30 min at 37°C in 3.0 mM Krebs-Ringer solution (pH 7.4, 0.1% BSA, 0.1 mM HEPES) followed by 30 min at 37°C in 17 mM Krebs-Ringer solution. Supernatants from both conditions were kept for insulin measurement. Insulin was measured using the Sensitive Insulin RIA kit (Linco Research).

Body composition: Body composition of the mice including fat and lean mass was determined by quantitative magnetic resonance imaging (MRI) using EchoMRI (Echo Medical Systems, Houston, TX, USA).
OGTT: Mice were fasted for 12 h from 9 pm with free access to water. Glucose (1.5 g/kg body weight) was administered orally. Concentrations of blood glucose were measured with a handheld glucometer (Ascensia Contour Glucometer, Bayer) using blood obtained from the punctures on the tail along a time course before and after glucose load. At time points of -60 and 15 min, blood samples were collected from the orbital sinus for insulin analysis (Sensitive Insulin RIA kit, Linco Research).