| Assay Method Information | |
| | Fluorescence Resonant Energy Transfer (FRET) Helicase Assay |
| Description: | The enzyme was assayed the DNA helicase activity by using DNA substrate (annealed BHQ1-labeled release strand and FITC-labeled template strand) in the presence of test compound. The substrate was monitored by the fluorescence quenching of FITC. The assay mixture was added to a 384-well microplate and monitored continuously at an excitation of 485 nm and an emission of 518 nm using a Fluoroskan Ascent microplate fluorescence reader (LabSystems, VA). The original data were normalized by the formula of relative helicase activity (%). When the inhibition of helicase activity was more than 50% at a concentration of 100 uM, the IC50 values were determined by curve fitting using SigmaPlot2000 of logistic with three parameters. |
| Affinity data for this assay | |
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