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2.4K Views
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08:11 min
November 11th, 2022
DOI :
10.3791/64805-v
Chapters
0:04
Introduction
1:04
Cell Culture
1:41
Fabrication of Micropipettes
2:16
Instrument Setup
2:37
IKv Parameter Setting
3:16
Whole‐Cell Patch‐Clamp Recording of the IKv in Voltage‐Clamp Mode
6:43
Results: IKv in H9c2 Cells
7:37
Conclusion
Transcript
该协议成功记录了乌头碱对H9C2细胞IKv的影响。从离子通道的角度推进药物心脏毒性机制研究的新阶段。全细胞膜片钳技术是离子通道研究的金标准,具有高灵敏度、高精度和高特异性。
全细胞膜片钳技术可用于探索并列门控多系统疾病(如心血管系统和神经系统)的特定离子通道的药理学或毒性机制。首先,一旦H9C2培养皿汇合80%,用0.25%胰蛋白酶消化细胞30秒。用普通培养基或含有药物的培养基在35毫米的培养皿中用玻璃板在37摄氏度下培养24小时,在5%二氧化碳气氛和70至80%相对湿度下培养
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Summary
本协议描述了一种使用全细胞膜片钳技术实时和动态采集H9c2心肌细胞中的电压门控钾(Kv)通道电流的有效方法。
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