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Analysis of Glucagon (Human, Mouse, Rat, Non-human primate, Canine) by ELISA (CAT#: STEM-MB-0863-LGZ)

Introduction

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 mini-glucagon, 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.




Principle

Enzyme-linked immunosorbent assay (ELISA) is an enzyme-labeled solid phase immunoassay technique. Its basic principle is to bind the antigen (or antibody) to the solid phase carrier, and the antigen (or antibody) and a certain enzyme link to enzyme labeled antigen (or antibody). During detection, the sample to be tested and the enzymic antigen (or antibody) react with the antigen (or antibody) on the solid phase carrier according to certain procedures, and then remove the unreacted part by washing method. After adding the substrate, the substrate is catalyzed by the enzyme on the solid phase carrier to produce colored substances. Through qualitative or quantitative detection of the amount of colored products, the content of the substance to be measured in the sample can be determined.

Applications

Glucagon measurements are used in the diagnosis and treatment of patients with various disorders of carbohydrate metabolism, including diabetes mellitus, hypoglycemia, and hyperglycemia.

Procedure

1. Add standards or samples to each well and incubate.
2. Pour off the liquid in the well, biotinylated antibody working solution and incubate.
3. Add enzyme conjugate working solution and incubate.
4. Add substrate TMB and incubate.
5. Add stop solution and measure OD value.
6. Calculation of results.

Materials

• Sample Type: Serum, plasma or supernatant

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