Contents of Kit
1. Capture Antibody (yellow): 0.5 ml of purified monoclonal anti-PC antibody for coating plates.
2. Detecting Antibody (red): 0.5 ml of peroxidase conjugated polyclonal anti-PC antibody for detection of captured PC.
Note: Reagents are sufficient for at least 5×96 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.
Protein C (PC) is a vitamin K-dependent glycoprotein produced in the liver. The concentration of PC in plasma is ~4 µg/ml (~60 nM). A deficiency of Protein C (quantitative or qualitative) is a risk factor for vascular thrombosis. Protein C is expressed as a two-chain molecule with a molecular weight of 62 kDa. The light chain (21 kDa) of PC consists of two EGF-like domains and an amino-terminal domain containing one hydroxyaspartic acid and 11 γ-carboxyglutamic acid (gla) residues. These residues allow PC to bind to membranes that contain acidic phospholipids in a calcium dependent manner. The heavy chain of PC (41 kDa) consists of the catalytic domain and an activation peptide. Activation of Protein C results from cleavage at residue Arg12 in the heavy chain by a complex of thrombin and a cell surface cofactor thrombomodulin. The activation of PC is associated with the release of a small activation peptide (2-3 kDa, called Protein C peptide, or PCP) from the N-terminal of the heavy chain. Activated Protein C (APC) is a serine protease with anticoagulant activity. APC, in complex with a phospholipid membrane, calcium and the Protein S cofactor, exhibits anticoagulant activity through the proteolytic inactivation of coagulation cofactors Va and VIIIa. The primary inhibitor of APC activity in plasma is Protein C Inhibitor (PCI, also called Plasminogen Activator Inhibitor-3, PAI-3) and to a lesser extent by α1 Antitrypsin and α2 macroglobulin. The inhibitory activity of PCI is stimulated approximately 10 fold by heparin.