TRAb clone ELISA Kit
TRAb clone ELISA Kit Developed and Manufactured by Medipan
Size: 1×96 wells
Sensitivity: 1.08 IU/l
Dynamic Range: 0.1 – 40 IU/l (WHO 08/204)
Incubation Time: 3 hours
Sample Type: Serum
Sample Size: 100 µL
Alternative Names: TSH Receptor Auto Antibody ELISA, Human TRAb clone ELISA
For Research Use Only
Controls Included
Assay Principle
The TRAb clone ELISA Kit is a competitive enzyme immunoassay with a step by step incubation. During the first incubation the TSH receptor antibodies of the patient samples and calibrators bind to the immobilized human recombinant receptor on the solid phase of the microtiter plate. Following an incubation period of 120 min, unbound antibodies are separated from the solid-phase immune complexes by a washing step. In a second incubation step of 20 min a biotinylated tracer antibody binds to the free epitopes of the receptor. The absence of serum autoantibodies against TSH receptor results in a complete saturation of the provided receptor by the biotinylated antibody. The presence of serum autoantibodies (TRAb) decreases the amount of biotinylated antibody bound. Unbound biotin complexes are removed by a washing step.
The complexed biotin reacts specifically with streptavidin horseradish peroxidase (HRP) conjugate. After the incubation period of 20 min, excessive conjugate is separated from the solid-phase immune complexes by the following washing step. Horseradish peroxidase converts the colorless substrate solution of 3,3’,5,5’-tetramethylbenzidine (TMB) added into a blue product. This enzyme reaction is stopped by dispensing an acidic solution (H2SO4) into the wells after 20 min at room temperature turning the solution from blue to yellow. The optical density (OD) of the solution at 450 nm is indirectly proportional to the amount of specific antibodies bound. The standard curve is established by plotting the concentrations of the antibodies of the standards (x-axis) and their corresponding OD values (y-axis) measured. The concentration of antibodies of the specimen is directly read off the standard curve.
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