Description
Principle of the Assay::The Human Glucagon ELISA (Enzyme-Linked Immunosorbent Assay) kit is an in vitro enzyme-linked immunosorbent assay for the quantitative measurement of Human Glucagon in Cell Culture Supernatants, Serum, Plasma. This assay employs an antibody specific for Human Glucagon coated on a 96-well plate. Standards and samples are pipetted into the wells and Glucagon present in a sample is bound to the wells by the immobilized antibody. The wells are washed and biotinylated anti-Human Glucagon antibody is added. After washing away unbound biotinylated antibody, HRP-conjugated streptavidin is pipetted to the wells. The wells are again washed, a TMB substrate solution is added to the wells and color develops in proportion to the amount of Glucagon bound. The Stop Solution changes the color from blue to yellow, and the intensity of the color is measured at 450 nm.
Background/Introduction: Glucagon is a 29 amino acid peptide produced by the pancreas that plays a critical role in glucose metabolism and homeostasis. The Glucagon precursor mRNA is expressed by alpha cells of the pancreas, L cells of the intestine, and in the brain. Only the pancreatic a-cells express the prohormone convertase PC2, also called PCSK2, which is required to produce Glucagon. Intestinal L cells instead express the prohormone convertase PC1, which processes the precursor to the Glucagon-overlapping peptides glicentin and oxyntomodulin. L cells also produce two Glucagon-like peptides, GLP-1 and GLP-2 that are derived from the same Glucagon precursor and influence glucose metabolism, but do not share any common sequence with Glucagon