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DOI: 10.3791/50227-v
Please note that some of the translations on this page are AI generated. Click here for the English version.
This article describes a method for preparing giant unilamellar vesicles that maintain the functional integrity of membrane proteins. The process involves depositing lipids onto a conductive surface, evaporating the solvent, rehydrating the lipid film, and applying an oscillating voltage to generate the vesicles.
重构功能膜蛋白组成界定成巨脂质体是一个功能强大的方法,结合膜片钳电。然而,传统的巨头脂质体产量可能与蛋白质的稳定性不相容。我们描述生产从单纯的脂质的巨脂质体或小脂质体含有离子通道的协议。
该程序的总体目标是以与掺入囊泡膜的膜蛋白的功能完整性相容的方式制备巨大的单层囊泡。这是通过首先将脂质或小脂质体沉积到导电表面(如 ITO 涂层载玻片)来实现的。第二步是以受控方式蒸发溶剂,以在导电表面上获得均匀的脂质膜。
接下来,在合适的缓冲液中小心地再水化脂质膜并准备电形成室。最后一步是在脂质膜上施加振荡电压,以产生巨大的 unal Miller 囊泡。最终,将对囊泡进行成像以确定它们的大小和质量,并与使用经典技术形成的囊泡进行比较。
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