Description(s):
GAD65 is one of the two isoforms of glutamate decarboxylase (GAD), an enzyme that catalyzes the decarboxylation of glutamate to produce gamma-aminobutyric acid (GABA). This enzyme is primarily expressed in the brain but can also be found in the insulin-producing beta cells of the pancreas. Although the function of GAD65 in the pancreas is not well understood, it is believed to help modulate the secretion of glucagon in alpha cells. GAD65 is a target of autoantibodies in people who later develop type 1 diabetes or latent autoimmune diabetes. Because of this, GAD autoantibodies are often used as a biomarker for these conditions.
Principle
The Mouse Anti-GAD65 ELISA Kit detects anti-GAD65 antibodies in mouse serum. This kit utilizes a plate coated with GAD65 to immobilize the autoantibodies of interest. Anti-mouse IgG antibody conjugated to horseradish peroxidase (HRP) is used to label the GAD65-bound antibodies, and the antibodies are detected by adding the HRP substrate, TMB, which forms a blue color in the presence of HRP. The color reaction is then terminated with Stop Solution, which causes the blue color to change to yellow. The autoimmune antibody concentration in each well is directly proportional to its color intensity and can be quantified by measuring its optical density at 450 nm (OD450) in a microplate reader.
