Description
Intended Uses: This human prothrombin antigen assay is intended for the quantitative determination of total prothrombin antigen in human plasma. For research use only.
Principle of the Assay: Human prothrombin will bind to the capture antibody coated on the microtiter plate. Prothrombin, thrombin, and thrombin-antithrombin complex will react with the antibody on the plate. After appropriate washing steps, anti-human prothrombin primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with the secondary antibody. Following an additional washing step, 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 human prothrombin. Color development is proportional to the concentration of total prothrombin in the samples.
Background: Prothrombin (aka Factor II) is a single-chain vitamin K-dependent 579 amino acid glycoprotein zymogen [1]. Prothrombin is proteolytically activated to thrombin by the prothrombinase enzyme complex in the coagulation cascade common pathway. The serine protease thrombin converts plasma fibrinogen to insoluble fibrin. Prothrombin levels are decreased by anticoagulant therapy, vitamin K deficiency and severe liver disease [2]. Elevated plasma prothrombin is associated with a single nucleotide change at position 20210 [3]