Source : Porcine Blood.
Thrombin enzyme (Activated Factor IIa) is an important clotting promoter that controls the transformation of soluble fibrinogen to insoluble active fibrin strands. Thrombin is a coagulation protein and a serine protease (EC 3.4.21.5) that catalyzes many coagulation-related reactions. Thrombin triggers factor-XI, factor-V, Factor-XIII and factor-VIII. Thrombin endorses platelet activation, using activation of protease-activated receptors on the platelet. As a result of its high proteolytic specificity, thrombin has become an important biochemical protein. The thrombin cleavage site (Leu-Val-Pro-Arg-Gly-Ser) is widely used in linker regions of recombinant fusion protein constructs. After the purification of the fusion protein, thrombin is used to cleave between the Arginine and Glycine residues of the cleavage site, efficiently removing the purification tag from the protein of interest with a high degree of specificity.
It is recommended to reconstitute the lyophilized Thrombin in sterile 18MΩ-cm H2O. One unit is defined as the amount of enzyme needed to cleave 1 mg of fusion protein in 16 hours to 95% completion at 20°C in a buffer containing 25mM Tris-HCl, pH 8.4, 150mM NaCl, and 2.5mM CaCl2.
