Human Factor V ELISA Kit（因子FV酶

Introduction
Factor V (FV) is an essential cofactor of the blood coagulation cascade and circulates in plasma as a large single-chain glycoprotein (330 kDa). The deduced amino acid sequence consists of 2224 amino acids inclusive of a 28-amino acid leader peptide (1). During coagulation, it is converted to the active cofactor FVa via limited proteolysis by thrombin, and spliced into a heavy chain (110 KDa) and a light chain (73 KDa) held together non-covalently by calcium (2). In the presence of a calcium ion and the phospholipid on cell surfaces, FVa and FXa form the prothrombinase complex which catalyzes the hydrolysis of prothrombin to thrombin (3). Thrombin in turn cleaves fibrinogen to fibrin which polymerizes to form a clot. FVa is readily inactivated by anticoagulant activated protein C (4). FV Leiden, a single amino acid mutation, renders FVa resistant to cleavage by activated protein C. It therefore over-produces thrombin and leads to excess clotting and hereditary thrombophilia disorder (5). Severe FV deficiency is associated with mild to severe bleeding diathesis (6).

Principle of the Assay
The AssayMax Human Factor V ELISA kit is designed for detection of human factor V in plasma, urine, milk, and cell culture supernatants. This assay employs a quantitative sandwich enzyme immunoassay technique that measures factor V in less than 4 hours. A monoclonal antibody specific for factor V has been pre-coated onto a microplate. Factor V in standards and samples is sandwiched by the immobilized antibody and a biotinylated polyclonal antibody specific for factor V, which is recognized by a streptavidin-peroxidase conjugate. All unbound material is then washed away and a peroxidase enzyme substrate is added. The color development is stopped and the intensity of the color is measured.