Direct detection of membrane channels from gels using water-in-oil droplet bilayers

被引:75
作者
Heron, Andrew J. [1 ]
Thompson, James R. [1 ]
Mason, Amy E. [1 ]
Wallace, Mark I. [1 ]
机构
[1] Univ Oxford, Dept Chem, Chem Res Lab, Oxford OX1 3TA, England
基金
英国生物技术与生命科学研究理事会; 英国工程与自然科学研究理事会;
关键词
D O I
10.1021/ja075715h
中图分类号
O6 [化学];
学科分类号
0703 ;
摘要
We form planar lipid bilayers between an aqueous droplet and a hydrogel support immersed in a lipid-oil solution. By scanning the bilayer over the surface of an SDS - PAGE gel, we are able to directly detect membrane proteins from gels using single-channel recording. Using this technique, we are able to examine low levels of endogenous protein from cell extracts without the need for over-expression. We also use droplet bilayers to detect small molecules from hydrogels. The bilayers show enhanced stability compared to conventional planar lipid bilayers, and both bilayer size and position can be controlled during an experiment. Hydrogel scanning with droplet bilayers provides a new method for the discovery and characterization of ion channels with the potential for high-throughput screening.
引用
收藏
页码:16042 / 16047
页数:6
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