| Assay Method Information | |
| | Electrophysiological Assay |
| Description: | A capillary glass pipette (BF150-86-10, Sutter Instruments) was pulled into a recording electrode with a micropipette puller (P97, Sutter Instruments). The micromanipulator (MP285, Sutter Instruments) was manipulated under an inverted microscope (IX71, Olympus) to make the recording electrode contact the cell and negative pressure suction was applied to achieve a GQ seal. Then, rapid capacitance compensation was performed, and negative pressure was continued, cell membrane was broken by suction, and whole cell recording mode was formed. Then the slow capacitance was compensated and the membrane capacitance and series resistance were recorded without leakage compensation. When the whole cell recording current was stabilized, dosing was initiated, and the next concentration was detected after acting 5 minutes of each drug concentration. Multiple cells were independently repeated during the recording period. All electrophysiological assays were performed at room temperature. Specifically, 6 concentrations (measuring IC50) or 2 concentrations (preliminary screening) were set for each compound, and the percentage of inhibition of sodium channels was determined by calculating the relative percentage of peak current generated before and after each concentration compound treated cells, and the IC50 value or percentage of inhibition at a specific concentration was calculated using IGOR pro software. |
| Affinity data for this assay | |
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