AssayPro—Human Glucokinase /He

Introduction
Human Glucokinase (GCK), also known as hexokinase IV or D, is a 50 kDa monomeric protein of 465 amino acids (1 - 2). It is present in the liver, pancreas, small intestine, and brain. It plays important roles in glucose metabolism. In response to rising levels of glucose from eating, GCK activity increases rapidly. It catalyzes the transfer of phosphate from ATP to glucose to form glucose-6-phosphate, which is the first rate-limiting step of glycogen synthesis and glycolysis. By means of this reaction, it functions as a glucose sensor for insulin secretion in pancreatic β-cells and regulates glucose and glycogen production in the liver (3). Mutations of the GCK gene are associated with non-insulin-dependent diabetes mellitus (4), persistent hyperinsulinemic hypoglycemia of infancy (5), and maturity-onset diabetes of younger individuals (6). GCK is a drug target for developing anti-type 2 diabetic molecules.

Principle of the Assay
The AssayMax Human Glucokinase/ Hexokinase-4 ELISA (Enzyme-Linked Immunosorbent Assay) kit is designed for detection of human GCK in plasma, serum, and cell culture supernatants. This assay employs a quantitative sandwich enzyme immunoassay technique that measures human GCK in 4 hours. A polyclonal antibody specific for human GCK has been pre-coated onto a 96-well microplate with removable strips. GCK in standards and samples is sandwiched by the immobilized antibody and the biotinylated polyclonal antibody specific for GCK, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.