Porcine insulin elisa kit

Most of the tested plasma parameters were determined with an automatic biochemical analyzer (Indiko Clinical Chemistry Analyzer, Thermofisher). Insulin levels were assessed by Elisa (Mercodia Porcine Insulin ELISA kit)). After KOH and MgCl2 extraction, liver TG levels were measured using a commercially available kit (TRIGLYCERIDES, Thermo Fisher Diagnostics). After extraction using the lipid extraction kit from Biovision, liver total cholesterol levels were determined with the Total cholesterol and Cholesteryl Ester Colorimetric Assay Kit II (Biovision).

Blood was drawn from the tail vein of mice and measured once a week for glucose measurements. An intraperitoneal glucose tolerance test (IPGTT) was performed in recipient mice at 4 and 8 weeks post-transplantation. After 15 hours of fast, 2 g/kg glucose was administered to mice, and blood glucose was measured at 0, 30, 60, 90, and 120 minutes.
Recipient blood samples were collected from NPCCs transplanted mice at 4 and 8 weeks post-transplantation. Blood was spun at 13,000 rpm for 10 minutes and blood serum was obtained and stored at –20°C. Serum insulin contents were evaluated by porcine insulin ELISA kit (Mercodia, Uppsala, Sweden) and measured by ELISA Reader (BIO-TEK, Winooski, VT, USA).

Mouse serum was obtained by centrifugation of blood collected from the cheek of the mouse every 2 or 3 weeks after transplantation. Porcine insulin levels in the serum of device-transplanted mice were measured using a porcine insulin ELISA kit (Mercodia). The crossover rate between porcine insulin and mouse insulin is 0.3% or less.

To evaluate insulin secretion from porcine islet-encapsulating devices, we performed glucose-stimulated insulin secretion of the device before transplantation, of the device after retrieval from the first transplanted mice, and of the device after retrieval from relay-transplanted mice. After the devices were retrieved from the abdominal cavities of the mice, the thin films that formed around the devices were removed before the glucose-stimulated insulin secretion test was performed. The devices were sequentially incubated at 37°C in medium (Functionality/Viability solution, CMRL 1066 [-] glucose, Corning, NY) containing low glucose (2.8 mM) for 2 h after two 30-min washes in a low-glucose solution and then incubated in medium containing high glucose (25 mM) for 2 h, before finally being incubated in medium containing low glucose (2.8 mM) for 2 h after a 1-h wash. The secretion of porcine insulin from devices was measured using a porcine insulin ELISA kit (Mercodia). Data are representative of 18 independent experiments and values are expressed as mean ± SEM