Contents of Kit
1. Capture Antibody (yellow): 0.4 ml of affinity-purified polyclonal anti-FX antibody for coating plates.
2. Detecting Antibody (red): 0.4 ml of peroxidase conjugated polyclonal anti-FX antibody for detection of captured murine FX.
Note: Reagents are sufficient for at least 4×96 well plates using recommended protocols. Antibodies are supplied in a 50% (v/v) glycerol solution for storage at –10 to -20°C. Keep vials tightly capped. Do not store in frost-free freezers.
Factor X (F.X, Stuart Factor) is a vitamin K-dependent glycoprotein produced in the liver. The concentration of F.X in plasma is ~10 µg/ml (~170 nM). Factor X is expressed as a two-chain molecule with a molecular weight of 59 kDa. The light chain (17 kDa) of F.X contains a calcium-binding domain consisting of one hydroxyaspartic acid and 11 γ-carboxyglutamic acid (gla) residues. These residues allow F.X to bind to membranes that contain acidic phospholipids in a calcium dependent manner. This is followed by two EGF-like domains. The heavy chain of F.X (42 kDa) consists of the catalytic domain, carbohydrate and the activation peptide. Activation of F.X to the active enzyme (F.Xa) results from cleavage at residue Arg52 in the heavy chain of F.X by a complex of F.IXa, cofactor VIIIa, calcium and negatively charged phospholipid surface (the tenase complex), or by the F.VIIa-tissue factor complex. Both activation pathways result in the release of the activation peptide from the N-terminal of the heavy chain. The F.Xa generated is a serine protease responsible for the activation of prothrombin to thrombin in the presence of a phospholipid membrane, calcium and cofactor Va. The activity of F.Xa in plasma is inhibited by antithrombin (ATIII), α1 Antitrypsin, α2 macroglobulin and tissue factor pathway inhibitor (TFPI). The inhibitory activity of ATIII is stimulated approximately 1000-fold by heparin.