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XB-ART-57323
Methods Mol Biol 2021 Jan 01;2191:49-63. doi: 10.1007/978-1-0716-0830-2_4.
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Characterizing Channelrhodopsin Channel Properties Via Two-Electrode Voltage Clamp and Kinetic Modeling.

Prignano L , Herchenroder L , Dempski RE .


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Two-electrode voltage clamp (TEVC) is a preferred electrophysiological technique used to study gating kinetics and ion selectivity of light-activated channelrhodopsins (ChRs). The method uses two intracellular microelectrodes to hold, or clamp, the membrane potential at a specific value and measure the flow of ions across the plasma membrane. Here, we describe the use of TEVC and a simple solution exchange protocol to measure cation selectivity and analyze gating kinetics of the C1C2 chimera expressed in Xenopus laevis oocytes. Detailed instructions on how to process the collected data and interpret the results are also provided.

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