| Solution Information | help | |
| Enzyme: | Potassium voltage-gated channel subfamily H member 2 | |
| inhibitor: | BDBM65906 | |
| substrate: | n/a | |
| Solution Type: | Aqueous | |
| pH at Preparation: | n/a | |
| Temp. Prep.: | n/a | |
| Comments: | Assay overview: The purpose of this assay is to test compound effects on reducing dofetilide-mediated inhibition using manual patch clamp for hERG-specific compounds. This assay employs a HEK293 cell line that stably expresses hERG channels. Cells are treated with test compounds, followed by measurement of electrophysiological activity of hERG using manual patch clamp. As designed, compound effects on the hERG channel were measured in the presence of dofetilide). Protocol: 1. Whole cell currents were generated form a holding potential of -80mV with a single step to +10 mV for 2 sec. followed by a tail pulse at -50 mV for 2 sec. The protocol was repeated every 10 sec. 2. Cell line used for patch clamp analysis was HEK cells stably expressing HERG kindly provided by Craig T. January (University of Wisconsin). This cell line is described in Zhou Z et al. Properties of HERG channels stably expressed in HEK 293 cells studied at physiological temperature. (Biophys J 74:230-241.) 3. The | |
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