| Assay Method Information | |
| | Inhibition of hERG Potassium Channel |
| Description: | Inhibition of hERG Potassium Channel by Prepared Compounds The final concentrations of test compounds were all prepared on the day of experiment and then dissolved in an extracellular fluid. The extracellular fluid (mM) was: NaCl, 137; KCl, 4; CaCl2, 1.8; MgCl2, 1; HEPES, 10; glucose 10; pH 7.4 (NaOH titration). All the test and control compound solutions contained 0.3% DMSO. HEK293 cells stably expressing the hERG ion channel were transferred to a perfusion chamber and perfusion was carried out using the extracellular fluid. A intracellular fluid was stored in small batches in a -80 C. freezer and thawed the day of experiment. The intracellular fluid (mM) was: K Aspartate, 130; MgCl2, 5; EGTA 5; HEPES, 10; Tris-ATP 4; pH 7.2 (KOH titration). Electrodes were produced by pulling with PC-10 (Narishige, Japan). Whole-cell patch-clamp recording was carried out. Noise was filtered at one fifth of the sampling frequency. The cells were clamped at -80 mV, then depolarized to 40 mV with a 4 second lasting square wave, and hyperpolarized to -40 mV with a 2 second lasting square wave to give a hERG tail current. This procedure was repeated every 20 seconds. The hERG tail current was a pure hERG current. The maximum current induced by the second square wave was detected, and after it was stable, perfusion was carried out using the test compounds. After the reaction was stable, the blocking intensity was calculated. |
| Affinity data for this assay | |
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