| Assay Method Information | |
| | hERG Kd Inhibition Assay |
| Description: | Compounds were dissolved in DMSO to a stock concentration of 10 mM and tested in 10-dose IC50 mode, in triplicate, with a 3-fold serial dilution starting at 100 uM. Control compound E-4031 was tested in 10-dose IC50 mode with 3-fold serial dilution starting at 1 uM. The assay is based on the competition of fluorescently labeled Tracer binding to the membrane preparation containing 1x Predictor hERG Membrane with 1 nM Predictor hERG Tracer Red in a buffer with the composition of 25 mM Hepes, pH 7.5, 15 mM KCl, 1 mM MgCl2, 0.05% PF-127, and 1% DMSO. Compounds in DMSO were added into the membrane mixture by using sonication, the tracer was added and gently mixed in the dark. The fluorescence was measured after 4 hours incubation at room temperature. The measurement parameters are as follows: Ex=531 nm FP and Em=595 nm P and S. |
| Affinity data for this assay | |
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