Description
Principle of the assay: The test kit is a solid phase enzyme immunometric assay (ELISA) in the microplate format, designed for the quantitative measurement of canine TSH. The microplate is coated with a monoclonal antibody specific for canine TSH. Calibrators and samples are pipetted into the antibody coated microplate. Afterwards, a polyclonal horseradish peroxidase-labeled antibody is added. During a two hour incubation sandwich complexes consisting of the two antibodies and the canine TSH is formed. Non-reactive components are removed by a washing step. A chromogenic substrate, TMB (3,3',5,5'-Tetra-Methyl-Benzidine), is added to all wells. During a 30 minutes incubation, the substrate is converted to a colored end product (blue) by the fixed enzyme. Enzyme reaction is stopped by dispensing of hydrochloric acid as stop solution (change from blue to yellow). The color intensity is direct proportional to the concentration of canine TSH present in the sample. The optical density of the color solution is measured with a microplate reader at 450 nm. Bi-chromatic measurement with a 600 - 690 nm reference filter is recommended