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用于平面脂质双层的伪绘画/气泡技术。

Pseudo painting/air bubble technique for planar lipid bilayers.

作者信息

Braun Christian J, Baer Tom, Moroni Anna, Thiel Gerhard

机构信息

Membrane Biophysics, Technical University of Darmstadt, Schnittspahnstrasse 3, 64287 Darmstadt, Germany.

Membrane Biophysics, Technical University of Darmstadt, Schnittspahnstrasse 3, 64287 Darmstadt, Germany.

出版信息

J Neurosci Methods. 2014 Aug 15;233:13-7. doi: 10.1016/j.jneumeth.2014.05.031. Epub 2014 Jun 2.

Abstract

BACKGROUND

A functional reconstitution of channel proteins in planar lipid bilayers is still very versatile to study structure/function correlates under well-defined conditions at the single protein level.

NEW METHOD

In this study we present an improved planar lipid bilayer technique in which an air bubble is used for stabilizing unstable/leaky bilayers or for removing excess lipids. The bubble can also be used as a tool for reducing the number of channels in the bilayer with the goal of having only one active channel in the membrane.

RESULTS

Stable planar lipid bilayers are formed within seconds to minutes. In the case of multiple channel insertion the air bubble can be used to reduce the number of channels within minutes.

COMPARISON WITH EXISTING METHOD(S): The simple improvement of the classical folding technique guarantees a very fast creation of stable bilayers even with difficult phospholipids in a conventional vertical bilayer set-up; it requires no modifications of the existing set-up.

CONCLUSIONS

This technique is very easy to handle and guarantees successful single channel recordings for any kind of planar lipid bilayer experiment.

摘要

背景

在平面脂质双分子层中对通道蛋白进行功能重建,在定义明确的条件下于单蛋白水平研究结构/功能相关性方面仍然非常通用。

新方法

在本研究中,我们提出了一种改进的平面脂质双分子层技术,其中使用气泡来稳定不稳定/渗漏的双分子层或去除多余的脂质。该气泡还可作为一种工具,用于减少双分子层中的通道数量,目标是使膜中仅存在一个活性通道。

结果

在数秒到数分钟内即可形成稳定的平面脂质双分子层。在多通道插入的情况下,气泡可用于在数分钟内减少通道数量。

与现有方法的比较

经典折叠技术的简单改进确保了即使在传统垂直双分子层设置中使用难处理的磷脂,也能非常快速地创建稳定的双分子层;无需对现有设置进行修改。

结论

该技术易于操作,可确保任何类型的平面脂质双分子层实验成功进行单通道记录。

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