Glutamic Acid Decarboxylase Autoantibody (GAD) ELISA Assay Kit
The Glutamic Acid Decarboxylase Autoantibody (GAD) ELISA Assay Kit is For Research Use Only
Sizes: 1 x 96 wells
Sensitivity: 0.8 AU/mL
Dynamic Range: 5 – 2000 U/ml
Sample Size: 25 uL
Incubation Time: 2 hours 35 minutes
Sample Type: Serum
Controls Not Included
Assay Background
Glutamic acid decarboxylase (GAD) autoantibodies are found in 70% to 80% of individuals with new-onset type 1 diabetes, making it the most frequent autoantibody in autoimmune diabetes. GAD autoantibodies can be detected in serum for many years post diagnosis, and high concentrations of GAD autoantibodies have been considered as a marker of faster β-cell exhaustion in these patients. Furthermore, GAD autoantibodies in non-diabetic individuals predicts the later development of type 1 diabetes. Besides autoimmune diabetes, GAD autoantibodies also exist in Stiff Man Syndrome, autoimmune poly-endocrinopathies, and some of Grave’s Disease patients.
Assay Principle
In this GADA ELISA, recombinant GAD protein coated onto plate wells can specifically recognize the GAD autoantibodies in human sera and calibrators. After a 1-hour incubation, GAD autoantibodies are captured by immobilized GAD protein while the unbound components were discarded and washed away. Afterwards, biotinylated GAD protein (GAD Biotin) is added for another round of incubation for 1 hour, wherein the GAD-Biotin detects GAD autoantibodies previously bound to GAD protein on the plate. After removal of nonspecific bindings, bound GAD-Biotin is revealed by addition of streptavidin horseradish peroxidase (STV-HRP), which specifically binds with biotin, followed by the substrate 3,3′,5,5′-Tetramethylbenzidine (TMB), which results in formation of a blue color. Color reaction will be further stopped by 2M H2SO4, transforming the blue color to yellow signals. The absorbance of yellow reaction mixture is measured by plate reader at 450nm and 405nm. The higher the reading is, the higher concentration of GAD autoantibodies. Low concentration of GAD autoantibodies (<18 u/ml) is recommended to be read off the 450nm calibration curve, while high value of GAD Glutamic Acid Decarboxylase (GAD) ELISA Assay Kit autoantibodies to be read off 405nm standard curve. The measuring interval is 5-2000U/ml (units are NIBSC 97/550)
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