| Assay Method Information | |
| | In Vitro Assay |
| Description: | Thrombin: In the assay for measuring the inhibition of human thrombin activity buffer (10 mM HEPES buffer, pH 7.4, 2 mM CaCl2) is used. The IC50 is measured by selecting the appropriate wells of a Greiner 384 microtiter plate. 50 μL the assay buffer containing human thrombin (Sigma; T8885), of which the final concentration is 0.05 NIH units/ml; 2 μL the assay buffer containing 2% (V/V) DMSO (uninhibited control) or various concentrations of the compounds to be tested diluted in the assay buffer containing 2% (V/V) DMSO; and add 48 μL the assay buffer containing substrate S-2238 (Chromogenix; Chemical Formula: H-D-Phe-Pip-Arg-pNA*2HCl), the final concentration is 30 μM. The assay is performed by pre-incubating the tested compounds with enzyme for 10 minutes. Then the assay is initiated by adding substrate to obtain a final volume of 100 μL. |
| Affinity data for this assay | |
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