| Assay Method Information | |
| | SyncroPatch 384 (Nanion) High Throughput Electrophysiology |
| Description: | Human Kv7.2, Kv7.4 or Kv7.5/7.3 channels were evaluated using a voltage protocol in which cells were voltage-clamped at a holding potential of -60 mV. Potassium currents were continuously activated with a series of three voltage steps to -30 mV for 3 seconds, 40 mV for 1 second and -90 mV for 4 seconds with 12 seconds between successive voltage sweeps. Potassium currents were measured from the -90 mV repolarizing step. Baseline current was assessed for 3.5 minutes prior to the addition of 5.6 uM zinc pyrithione (1 uM for Kv7.4). Kv7.2, Kv7.4 or Kv7.5/7.3 current in the presence of zinc pyrithione was acquired over five minutes to allow channels to reach steady state activity prior to addition of test agents. Channel activity was monitored for three minutes preceding the addition of 30 uM ML-213 (3 minutes) to achieve maximum activation. 150 mM TEA with 10 uM XE-991 was applied for 2 minutes to measure the leak current during maximum inhibition of Kv7.2, Kv7.4 or Kv7.5/7.3 channels. |
| Affinity data for this assay | |
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