Tyrosine Phosphatase-related Islet Antigen 2 AutoAb (IA2A) ELISA Assay Kit
The Tyrosine Phosphatase-related Islet Antigen 2 AutoAb (IA2A) ELISA Assay Kit is For Research Use Only
Sizes: 1 x 96 wells
Sensitivity: 0.8 AU/mL
Dynamic Range: 7.5 – 4000 U/mL
Sample Size: 25 uL
Incubation Time: 16 hours – 21 hours 35 minutes
Sample Type: Serum
Controls Not Included
Assay Background
The prevalence of autoantibodies against protein tyrosine phosphatase 2 (IA-2/ICA512) is 55-75% in newly onset Type I diabetes (T1D) patients. T1D patients who process autoantibodies against IA-2 have a higher risk of faster T1D development. IA-2 autoantibodies detection is able to increase the detection rate of T1D to 94%. Besides T1D, IA-2 autoantibodies also exist in autoimmune thyroid diseases (AITD).
Assay Principle
In the Tyrosine Phosphatase-related Islet Antigen 2 AutoAb (IA2A) ELISA, recombinant IA-2 protein coated onto plate wells can specifically recognize the IA-2 autoantibodies in human sera and calibrators. After a 16–20-hour incubation at 4℃, IA-2 autoantibodies are captured by immobilized IA-2 protein while the excess or unbound components were discarded and washed away. Afterwards, biotinylated IA-2 protein (IA2-Biotin) is added for another round of incubation for 1 hour at 4℃ , wherein the IA2-Biotin detects IA-2 autoantibodies previously bound to IA-2 protein on the plate. After removal of nonspecific bindings, bound IA2-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 IA-2 autoantibodies. Low concentration of IA-2 autoantibodies (<15 u/ml) is recommended to be read off the 450nm calibration curve, while high value of IA-2 autoantibodies to be read off 405nm standard curve. The measuring interval is 5-2000U/ml (units are NIBSC 97/550)
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