Microvette cb 300 k2 edta tubes

Plasma insulin from the neonates was measured on post-natal day 4 using Ultrasensitive mouse insulin ELISA kit (Catalog no. 10-1249-01). In adult mice, plasma insulin measurements were conducted in mice fasted for ~4 hours followed by intraperitoneal injection of 2mg dextrose/g body weight at 0-, 15-, and 30-min post injection. Tail blood samples were collected in Microvette^® CB 300 K2 EDTA tubes (Catalog no. 16.444.100, Sarstedt) at the indicated time points and plasma insulin were measured using mouse ultrasensitive insulin ELISA kit (Mercodia Inc., Sweden). For in vitro insulin and glucagon secretion assays, islets were isolated from mice fed a standard chow diet and were incubated over-night in RPMI supplemented with 0.5mg/mL BSA. On the following day, islets were equilibrated in DMEM containing 0.5 mg/mL BSA, 0.5 mM CaCl₂ and 10.0 mM HEPES (DMEM*) supplemented with 10% FBS and 5.5 mM glucose for 1 h at 37°C, 5% CO₂. 20 islets/well were picked into 400 uL DMEM* without FBS at glucose concentrations specified in the figures in 24-well plate(s) and insulin or glucagon secretion was measured over 1 h at 37°C and stored at −20°C until analysis. Insulin secretion was measured using mouse insulin ELISA kit (Catalog no. 10-1247-01, Mercodia Inc, Sweden) and glucagon secretion was measured using the mouse glucagon ELISA kit (and 10-1281-01, Mercodia Inc, Sweden).

Plasma insulin from the neonates was measured on P4 using Ultrasensitive mouse insulin ELISA kits (Catalog no. 10-1249-01). In adult mice, plasma insulin measurements were conducted in mice fasted for ~4 hr followed by intraperitoneal injection of 2 mg dextrose/g body weight at 0, 15, and 30 min post injection. Tail blood samples were collected in Microvette CB 300 K2 EDTA tubes (Catalog no. 16.444.100, Sarstedt) at the indicated time points and plasma insulin were measured using mouse ultrasensitive insulin ELISA kits (Mercodia Inc, Sweden). For in vitro insulin and glucagon secretion assays, islets were isolated from mice fed a standard chow diet and were incubated overnight in RPMI supplemented with 0.5 mg/ml BSA. On the following day, islets were equilibrated in DMEM containing 0.5 mg/ml BSA, 0.5 mM CaCl₂ and 10.0 mM HEPES (DMEM*) supplemented with 10% FBS and 5.5 mM glucose for 1 hr at 37°C, 5% CO₂. 20 islets/well were picked into 400 µl DMEM* without FBS at glucose concentrations specified in the figures in 24-well plate(s) and insulin or glucagon secretion was measured over 1 hr at 37°C and stored at –20°C until analysis. Insulin secretion was measured using mouse insulin ELISA kits (Catalog no. 10-1247-01, Mercodia Inc, Sweden) and glucagon secretion was measured using mouse glucagon ELISA kits (Catalog no. 10-1281-01, Mercodia Inc, Sweden).