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Application: Screen small molecule inhibitors in high throughput screening (HTS) applications.
Background: 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.
Description: The Factor Xa Inhibitor Screening Assay is a colorimetric assay designed to measure the activity of human Factor Xa for screening and profiling applications. The assay kit comes in a convenient 96-well format or 384-well format and contains enough purified human Factor Xa, a chromogenic substrate, and PR-02 buffer for 100 or 400 reactions.To determine the effect of an inhibitor on Factor Xa activity, the enzyme should be preincubated with or without the test inhibitor prior to adding the chromogenic substrate to the reaction. The assay was functionally validated using Rivaroxaban, a potent inhibitor of Factor Xa.Upon proteolysis, Factor Xa cleaves the chromogenic substrate at the C-terminal end releasing p-nitroanilid (pNA), which produces a yellow color that is measurable photometrically at λ=405 nm. The increase in color is proportional to Factor Xa activity.
Storage Stability: This assay kit will perform optimally for up to 6 months from date of receipt when the materials are stored as directed.
Warnings: Avoid freeze/thaw cycles
Biosafety Level: Not applicable (BSL-1)
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