Contents of Kit
1. Capture Antibody (yellow): 0.4 ml of polyclonal affinity purified anti-PK antibody for coating plates.
2. Detecting Antibody (red): 0.4 ml of peroxidase-conjugated polyclonal anti-PK antibody for detection of captured PK.
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.
Prekallikrein (PK), previously known as Fletcher Factor, is a single chain glycoprotein produced in the liver. The plasma concentration of PK is 50 μg/ml (550 nM), approximately 75% of which circulates in complex with high molecular weight kininogen (HK) and the remainder as free PK. Plasma PK is heterogeneous in both mass and charge due to variable degrees of glycosylation. Approximately 90% of plasma PK has an apparent molecular weight of 88 kDa as determined by SDS-PAGE and the remaining 10% has an apparent mass of 85 kDa. The catalytic site resides in the light chain. The heavy chain of PK contains four apple-domain structures similar to those found in F.XI and these are required for binding of PK to HK. PK is the zymogen form of the enzyme kallikrein, which is involved in the proteolysis of kininogens with subsequent release of bradykinin, a potent vasodilator. PK participates in the contact phase of coagulation as a substrate for surface-bound activated factor XII (F.XIIa) in the presence of the cofactor HK. As PK and factor XI (F.XI) both circulate in complex with HK, both are localized to activating surfaces through their respective binding to HK. Limited proteolysis of PK by F.XIIa generates kallikrein, a two-chain serine protease that initiates the reciprocal activation of PK and F.XI. Kallikrein activity in plasma is regulated predominantly by C1-Inhibitor and α2 macroglobulin, with relatively minor contributions by Protein C Inhibitor, α2 Antiplasmin, and antithrombin.