Contents of Kit
1. Capture Antibody (yellow): 0.5 ml of polyclonal affinity purified anti-FXI antibody for coating plates.
2. Detecting Antibody (red): 0.5 ml of peroxidase conjugated polyclonal anti-FXI antibody for detection of captured FXI.
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.
Factor XI (F.XI, plasma thromboplastin antecedent) is a coagulation protein produced in the liver that circulates in plasma at approximately 5 µg/mL (30 nM). The mass of F.XI is 160 kDa as determined by SDS-PAGE under non-reducing conditions and 80 kDa upon reduction. F.XI consists of two identical 80 kDa subunits linked by disulphide bonds. Each subunit consists of a tandem repeat of four apple domains followed by a serine protease catalytic domain. Cleavage of F.XI by activated factor XII or thrombin converts each subunit into a two-chain form and generates two active sites per F.XIa molecule. The mass of F.XIa is 160 kDa unreduced, but upon reduction F.XIa migrates as a heavy chain of 50 kDa and a light chain of 30 kDa. The catalytic site of F.XIa resides in the light chain. In plasma, F.XI or F.XIa circulates in non-covalent 1:1 complex with high molecular weight kininogen, which acts as a cofactor in the activation of F.XI by activated factor XII. The activity of F.XIa is regulated by platelets and by several proteinase inhibitors including, in order of decreasing importance, C1-inhibitor, α2 Antiplasmin, α1 Antitrypsin and antithrombin. Heparin has relatively little effect on the rate of inhibition of F.XIa by antithrombin. The only known natural substrate for activated F.XI (F.XIa) is factor IX (Christmas factor) and the only cofactor required for this reaction is ionized calcium.