Screening Assay Protocols Targeting the Nav1.7 Channel Using Qube High-Throughput Automated Patch-Clamp System

Q2 Pharmacology, Toxicology and Pharmaceutics Current Protocols in Pharmacology Pub Date : 2020-04-13 DOI:10.1002/cpph.74

Binbin Qian, Sung Hoon Park, Weifeng Yu

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引用次数: 6

Abstract

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 V_half, 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 V_half 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. © 2020 by John Wiley & Sons, Inc.

Basic Protocol 1: Comparing V_half measurements using offline calculation and a new adaptive online estimation

Basic Protocol 2: Concentration response experiments using a state-dependent protocol

Basic Protocol 3: Concentration response experiments using a use-dependent protocol

Support Protocol: Cell culture protocol

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使用Qube高通量自动膜片钳系统筛选Nav1.7通道的检测方案

本文介绍了在Qube自动膜片钳系统上设置筛选协议和记录Nav1.7数据的基本过程。提供了三个协议以及逐步的细节。首先，我们描述了一个估计Vhalf的协议，即一半通道失活时的电压，使用传统的稳态失活测量以及一种新的自适应在线估计。其次，我们建立了一个状态依赖的协议，使用自适应在线Vhalf测量来获得已知参考阻断剂的浓度响应曲线(CRC)。最后，我们介绍了一个与使用相关的协议。在我们的研究中，样品参照物显示出良好的状态和用途依赖的Nav1.7抑制作用。©2020 by John Wiley &基本协议1:使用离线计算和新的自适应在线估计比较Vhalf测量基本协议2:使用状态依赖协议的浓度响应实验基本协议3:使用使用依赖协议的浓度响应实验支持协议:细胞培养协议

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Current Protocols in Pharmacology Pharmacology, Toxicology and Pharmaceutics-Pharmacology

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