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Assay Method Information |
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| | Human Thrombin Assay |
| Description: | The inhibitory activity on human thrombin was determined by using a buffer (10 mM HEPES buffer, pH 7.4, 2 mM CaCl2). Appropriate wells in the Greiner 384 microtiter plate were used to determine IC50. A buffer contained 50 μL human thrombin (Sigma; T8885), the final concentration of which was 0.05 NIH unit/mL. A buffer containing 2 μL 2% (V/V) DMSO (control group which was free of inhibition) or various concentration of the compound to be tested was diluted in the buffer containing 2% (V/V) DMSO. A buffer containing 48 μL substrate S-2238 (Chromogenix; Chemical formula: H-D-Phe-Pip-Arg-pNA.2HCl) was added, the final concentration was 30 μM. In this assay, the compound to be tested and the enzyme were pre-incubated for 10 minutes, the substrate was added to give a final volume of 100 μL to start the assay. |
| Affinity data for this assay | |
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