Introduction:
Factor X, also known as EC3.4.21.6 or thrombokinase, is a serine endopeptidase involved in blood coagulation. Factor X needs to be activated into Factor Xa to cleave prothrombin into active thrombin. Deficiency in inactive and active Factor X results in blood coagulation disorders. Warfarin, a common anticoagulant used in the treatment of thrombosis, inhibits the conversion of Factor X to Xa, resulting in lower risk of stroke and thrombosis. Warfarin is used to treat pulmonary embolism and lower the risk of cardiovascular complications. The development of new Factor Xa inhibitors, also known as blood thinners, has been crucial in improving the quality of life of patients by bypassing the need for frequent blood tests.
Intended Use:
The KRISHZYME Factor Xa Inhibitor Screening Assay Kit colorimetric assay designed to measure the activity of human Factor Xa for screening and profiling applications.
Principle:
The assay is based on a chromogenic method in which Factor Xa cleaves a specific synthetic substrate to release p-nitroaniline (pNA), producing a yellow colour measurable at 405 nm. To evaluate inhibitory effects, Factor Xa is preincubated with or without a test inhibitor prior to substrate addition. In the absence of an inhibitor, active Factor Xa cleaves the substrate efficiently, resulting in increased colour intensity, whereas in the presence of an inhibitor, enzymatic activity is reduced, leading to decreased pNA release and lower absorbance. Thus, the intensity of the colour is directly proportional to Factor Xa activity and inversely proportional to the inhibitory effect of the test compound.
