This article describes the basic procedure for setting up the screening protocol and recording data for Nav1.7 on a Qube automated patch-clamp system. Three protocols along with step-by-step details are provided. First, we describe a protocol to estimate Vhalf , the voltage at which half of the channels are inactivated, using traditional steady-state inactivation measurement as well as a new adaptive online estimation. Second, we establish a state-dependent protocol using adaptive online Vhalf measurement to obtain a concentration response curve (CRC) on known reference blockers. Last, we introduce a use-dependent protocol. In our hands, the sample reference demonstrated good state- and use-dependent inhibition of Nav1.7.
This article describes the basic procedure for setting up the screening protocol and recording data for Nav1.7 on a Qube automated patch-clamp system. Three protocols along with step-by-step details are provided. First, we describe a protocol to estimate Vhalf , the voltage at which half of the channels are inactivated, using traditional steady-state inactivation measurement as well as a new adaptive online estimation. Second, we establish a state-dependent protocol using adaptive online Vhalf measurement to obtain a concentration response curve (CRC) on known reference blockers. Last, we introduce a use-dependent protocol. In our hands, the sample reference demonstrated good state- and use-dependent inhibition of Nav1.7.
Authors: Grant P Higerd-Rusli; Matthew Alsaloum; Sidharth Tyagi; Nivedita Sarveswaran; Mark Estacion; Elizabeth J Akin; Fadia B Dib-Hajj; Shujun Liu; Daniel Sosniak; Peng Zhao; Sulayman D Dib-Hajj; Stephen G Waxman Journal: J Neurosci Date: 2022-05-18 Impact factor: 6.709
Authors: Julie I R Labau; Matthew Alsaloum; Mark Estacion; Brian Tanaka; Fadia B Dib-Hajj; Giuseppe Lauria; Hubert J M Smeets; Catharina G Faber; Sulayman Dib-Hajj; Stephen G Waxman Journal: Front Pharmacol Date: 2021-12-21 Impact factor: 5.810