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Assay Method Information

Assay Name:  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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Last update November 1, 2007
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