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| | Prothrombin Assay |
| Description: | The activity of the compound to be tested against prothrombinase was determined by the production of thrombin. In summary, 12.5 μL human factor Xa was incubated in 10 mM HEPES buffer and pH 7.4, 2 mM CaCl2, the final concentration was 0.5 nM, and 12.5 μL human blood platelets (1×107 mL−1) was added at 37° C. After 10 mins, 25 μL prothrombin was added to initiate the reaction, the final concentration was 0.5 μM, a buffer containing 2 μL 2% (V/V) DMSO (control group which was free of inhibition) or various concentration of the compound was diluted in the buffer containing 2% (V/V) DMSO. After 20 minutes, 48 μL substrate S-2238 (Chromogenix) was added until the final concentration was 50 μM to determine thrombin activity. |
| Affinity data for this assay | |
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