Pancreatic islets of Langerhans were isolated from wild type and GPR40−/− mice (littermates) by collagenase digestion and discontinuous Ficoll gradient separation [14 (link)]. The islets were cultured overnight in RPMI-1640 medium with 11 mmol/l glucose to facilitate recovery from the isolation process. Insulin secretion was determined by a 1-h static incubation in Krebs-Ringer bicarbonate (KRB) buffer in a 96-well format as previously described [15 (link),16 (link)]. Briefly, islets were first preincubated in KRB medium with 2 mmol/l glucose for 30 min and were then transferred to a 96-well plate (three islets/well) and incubated with 200 μl of the KRB medium with 15 mmol/l glucose in the presence or absence of testing compounds for 60 min. The buffer was removed from the wells at the end of the incubation and assayed for insulin levels using the Insulin ELISA kit (ALPCO, Salem, NH) and glucagon level using glucagon ELISA kit (ALPCO, Salem, NH).
Glucagon elisa kit
Manufactured by ALPCO
Sourced in United States
The Glucagon ELISA kit is a quantitative immunoassay designed for the measurement of glucagon in biological samples such as plasma, serum, and cell culture supernatants. The kit uses a sandwich ELISA principle to detect and quantify glucagon levels.
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Lab products found in correlation
2 protocols using glucagon elisa kit
1
Pancreatic Islet Isolation and Insulin Secretion
2
Intravenous Glucose Tolerance Test in Mice
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After the animals were fasted for 8 h, their blood glucose levels were measured with a SureStep Blood Glucose meter (LifeScan, Milpitas, CA, USA). An intravenous glucose tolerance test (IVGTT) was performed by tail vein injection of 1 g/kg D-glucose (Sigma-Aldrich; Merck KGaA) on days 14 and 28. Plasma insulin levels were determined by using an ultrasensitive mouse insulin ELISA kit (cat. no. 80-INSMS-E01; ALPCO, Salem, NH, USA) and plasma glucagon levels were determined by using a glucagon ELISA kit (cat. no. 81518; Crystal Chem Inc., Downers Grove, IL, USA).
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