Rat mouse insulin elisa kit

For pancreatic islet isolation, ice-cold 0.45 mg/mL collagenase type V buffer (Sigma-Aldrich, St. Louis, MO) (in Hanks’ balanced salt solution without Ca²⁺) was perfused into the bile duct. The pancreas was immediately dissected out and incubated at 37°C for 10–11 min in a water bath for digestion, and islets were washed twice in Hanks’ balanced salt solution and then handpicked under a stereo microscope. Islets were incubated overnight in RPMI complete media supplemented with 10% FCS, 100 units/mL penicillin, and 100 μg/mL streptomycin (Thermo Fisher Scientific). For GSIS, islets were preincubated in Krebs-Ringer buffer solution (bicarbonate HEPES) containing 2% BSA and 2.8 mmol/L glucose for 1 h. Islets were then separated into groups of 10 and incubated with either 2 or 20 mmol/L glucose for 1 h, and media were collected for GSIS. Total insulin content was extracted from 10 islets in 0.2 N/80% acid ethanol. Insulin released into the medium and total insulin content were measured using a rat/mouse insulin ELISA kit (Crystal Chem) (25 (link)). Total proinsulin content was measured using a rat/mouse proinsulin ELISA kit (Mercodia).