Description
Intended Uses: This mouse prothrombin/thrombin total antigen assay is intended for the quantitative determination of total prothrombin and thrombin in mouse plasma, serum, tissue extracts, and cell culture media. For quantitation of mouse prothrombin only, refer to the Mouse Prothrombin Antigen Assay (cat # MPTKT). For research use only.
Principle of the Assay: Mouse prothrombin, thrombin, and thrombin-antithrombin complex will bind to the capture antibody coated on the microtiter plate. After appropriate washing steps, biotinylated primary antibody binds to the captured protein. Excess primary antibody is washed away and bound antibody is reacted with horseradish peroxidase conjugated streptavidin. 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 prothrombin. The amount of color development is directly proportional to the concentration of prothrombin in the sample.
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]