Thyrotropin Receptor Autoantibodies (TRAb) ELISA

Enzyme immunoassay for the quantitative determination of thyrotropin receptor autoantibodies (TRAb) in human serum.

INTENDED USE:The RSR TSH receptor (TSHR) autoantibody (TRAb) ELISA kit is intended for use by professional persons only for the quantitative determination of TRAb in human serum. Hyperthyroidism in Graves’ disease is due to the presence of autoantibodies to the TSHR and measurement of these autoantibodies can be useful in disease diagnosis and management.

GENERAL INFORMATION:In RSR’s TRAb ELISA, TRAb in patients’ sera, calibrators and controls are allowed to interact with TSHR coated onto ELISA plate wells. After a 2 hour incubation, the samples are discarded leaving TRAb bound to the immobilised TSHR. TSH-Biotin is added in a 2nd incubation step, where it interacts with immobilised TSHR which have not been blocked by the bound TRAb from patient sera, calibrators or controls. The amount of TSH-Biotin bound to the plate is then determined in a 3rd incubation step by addition of Streptavidin Peroxidase (SA-POD), which binds specifically to Biotin. Excess unbound SAPOD is then discarded and the addition of the peroxidase substrate 3,3’,5,5’–tetramethylbenzidine (TMB) results in the formation of a blue colour. This reaction is stopped by the addition of stop solution causing the well contents to turn from blue to yellow. The absorbance of the yellow reaction mixture at 450nm is then read using an ELISA plate reader. A lower absorbance indicates the presence of TRAb in a test sample as TRAb inhibits the binding of TSH-Biotin to TSHR coated plate wells. The measuring range is 1 – 40 IU/L (NIBSC 08/204).

KIT CHARACTERISTICS:
- Method: ELISA
- Tests: 96
- Incubation Time / Conditions: 2 h, 20 min, 20 min, 20 min
- Standard Range: 1 - 40 U/l
- Sensitivity analytical: 0.55 U/l
- Final Sample Volume: 75 µl
- Sample Type: serum
- Isotope / Substrate: TMB 450 nm
- Internal Controls: 2
- Regulatory Status: CE