Mouse/Rat Glucagon ELISA Assay Kit
The Mouse/Rat Glucagon ELISA Assay Kit was manufactured in Sweden by Mercodia
Size: 12 x 8 wells
Sensitivity: 1.5 pmol/L
Dynamic Range: 2 – 180 pmol/L (7 – 627 pm/mL)
Incubation Time: Overnight
Sample Type: Serum, EDTA plasma, cell-culture medium
Sample Size: 10 µL
Assay Background
Glucagon is a 29 amino acid polypeptide processed from proglucagon in pancreatic alpha cells. In intestinal L-cells proglucagon is cleaved into glicentin, corresponding to proglucagon residues no 1-69. Glicentin can further be processed into oxyntomodulin, corresponding to proglucagon residues no 33-69. These peptides are released simultaneously upon stimulation. Moreover, a fragment of glucagon corresponding to its C-terminal part (residues no 19-29), also designated miniglucagon, is reported to be present in the pancreas in low amounts compared to the total glucagon content. In general, glucagon has an effect opposite that of insulin, i.e. it raises blood glucose levels. It causes the liver to convert glycogen into glucose, which is then released into the blood stream. With longer stimulation, glucagon action in the liver results in a glucose-sparing activation of free fatty acid oxidation and production of ketones. During hypoglycaemia, glucagon secretion offers a protective feedback mechanism, defending the organism against damaging effects of glucose deficiency in the brain and nerves.
Assay Principle
The Mouse/Rat Glucagon ELISA Assay Kit is a solid phase two-site enzyme immunoassay. It is based on the direct sandwich technique in which two monoclonal antibodies are directed against separate antigenic determinants on the glucagon molecule. During incubation glucagon in the sample reacts with peroxidase-conjugated antiglucagon antibodies (clone E6A11K) and anti-glucagon antibodies (clone M5F9S) bound to microplate wells. A simple washing step removes unbound enzyme labelled antibody. The bound conjugate is detected by reaction with 3,3’,5,5’-tetramethylbenzidine (TMB). The reaction is stopped by adding acid to give a colorimetric endpoint that is read spectrophotometrically.
