| Assay Method Information | |
| | Enzyme Assay |
| Description: | The rate of hydrolysis of the substrates (0.3 mM) by trypsin (18 U/ml), thrombin (12.5 NIH U/ml), urokinase (62500 IU/ml) and elastase (8.4 U/ml) was monitored at410 nm in Tris-HCl buffer (50 mM, pH 8.0) without the presence of Ca2+ or other activators, at 37°C, pH 7.6. Data scanning time was set to the 1st and 61st minute after the reaction start. Rutin, esculin, phloridzin and their esters were initially solubilized at a concentration of 10 mM in dimethyl sulphoxide (DMSO) and subsequently incorporated in the reaction mixture to final concentrations in the range of 1-50 μM. Samples were run in triplicate in 96-well plates by MRX microplate reader. Each experiment was performed in triplicate. |
| Affinity data for this assay | |
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