{"title":"使用Qube高通量自动膜片钳系统筛选Nav1.7通道的检测方案","authors":"Binbin Qian, Sung Hoon Park, Weifeng Yu","doi":"10.1002/cpph.74","DOIUrl":null,"url":null,"abstract":"<p>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 <i>V</i><sub>half</sub>, 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 <i>V</i><sub>half</sub> 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.</p><p><b>Basic Protocol 1</b>: Comparing <i>V</i><sub>half</sub> measurements using offline calculation and a new adaptive online estimation</p><p><b>Basic Protocol 2</b>: Concentration response experiments using a state-dependent protocol</p><p><b>Basic Protocol 3</b>: Concentration response experiments using a use-dependent protocol</p><p><b>Support Protocol</b>: Cell culture protocol</p>","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"89 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-04-13","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.74","citationCount":"6","resultStr":"{\"title\":\"Screening Assay Protocols Targeting the Nav1.7 Channel Using Qube High-Throughput Automated Patch-Clamp System\",\"authors\":\"Binbin Qian, Sung Hoon Park, Weifeng Yu\",\"doi\":\"10.1002/cpph.74\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>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 <i>V</i><sub>half</sub>, 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 <i>V</i><sub>half</sub> 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.</p><p><b>Basic Protocol 1</b>: Comparing <i>V</i><sub>half</sub> measurements using offline calculation and a new adaptive online estimation</p><p><b>Basic Protocol 2</b>: Concentration response experiments using a state-dependent protocol</p><p><b>Basic Protocol 3</b>: Concentration response experiments using a use-dependent protocol</p><p><b>Support Protocol</b>: Cell culture protocol</p>\",\"PeriodicalId\":10871,\"journal\":{\"name\":\"Current Protocols in Pharmacology\",\"volume\":\"89 1\",\"pages\":\"\"},\"PeriodicalIF\":0.0000,\"publicationDate\":\"2020-04-13\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"https://sci-hub-pdf.com/10.1002/cpph.74\",\"citationCount\":\"6\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Current Protocols in Pharmacology\",\"FirstCategoryId\":\"1085\",\"ListUrlMain\":\"https://onlinelibrary.wiley.com/doi/10.1002/cpph.74\",\"RegionNum\":0,\"RegionCategory\":null,\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"Pharmacology, Toxicology and Pharmaceutics\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpph.74","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Pharmacology, Toxicology and Pharmaceutics","Score":null,"Total":0}
引用次数: 6
Screening Assay Protocols Targeting the Nav1.7 Channel Using Qube High-Throughput Automated Patch-Clamp System
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. © 2020 by John Wiley & Sons, Inc.
Basic Protocol 1: Comparing Vhalf 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
求助内容：
应助结果提醒方式：
京公网安备 11010802042870号
