Description
Principle of the assay: Human TAT complex in samples will bind to the anti-human thrombin capture antibody coated on the microtiter plate. After appropriate washing steps, monoclonal anti-human antithrombin primary antibody binds to TAT complex captured on the plate. Excess antibody is washed away and bound monoclonal antibody is then reacted with the secondary antibody conjugated to horseradish peroxidase. TMB substrate is used for color development at 450nm. A standard calibration curve is prepared along with the samples to be measured using dilutions of TAT complex. Color development is proportional to the concentration of TAT complex in the samples. Free thrombin, prothrombin and antithrombin III will not be detected by this assay.
Background: Antithrombin III is a glycosylated plasma serine protease inhibitor that forms a stoichiometric complex with coagulation cascade enzymes [1]. Thrombin is a two-chain vitamin K-dependent glycosylated serine protease that is activated from prothrombin in the coagulation cascade [2]. Antithrombin III inhibits thrombin with heparin enhanced kinetics and forms a 1:1 covalent complex [3]