| Assay Method Information | |
| | TREK-1 Manual Patch Clamp (hMPC) assay |
| Description: | TBDCHO-K1 cells stably expressing human TREK-1 or HEK293 cells stably expressing human TREK-2 are plated on glass coverslips, and voltage clamped in the whole-cell configuration of the patch clamp technique. Cells were voltage clamped at a holding potential of −80 mV and the stepped to 0 mV for 500 msec. The voltage was subsequently ramped from −120 mV to +80 mV over a 500 msec duration. This step-ramp protocol was repeated every 10 sec. The bathing solution contained the following: 135 mM NaCl, 5 mM KCl, 2 mM CaCl2, 1 mM MgCl2, 5 mM D-Glucose, 10 mM HEPES, 10 mM sucrose (adjusted to pH 7.4 with NaOH, 300 mosmol/kg H2O). The pipette solution contained the following: 135 mM KCl, 2 mM MgCl2, 1 mM EGTA, 10 mM HEPES, 2 mM Na 2 ATP (adjusted to pH 7.35 with KOH, 285 mosmol/kg H2O). |
| Affinity data for this assay | |
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