Direct detection of membrane channels from gels using water-in-oil droplet bilayers.
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.
Chemistry Research Laboratory, Department of Chemistry, University of Oxford, Mansfield Road, Oxford OX1 3TA, UK.
Andrew J
AJ
Heron
James R
JR
Thompson
Amy E
AE
Mason
Mark I
MI
Wallace
D004566:Electrodes; D004591:Electrophoresis, Polyacrylamide Gel; D005782:Gels; D008051:Lipid Bilayers; D008565:Membrane Proteins / Q000737:chemistry*; D009821:Oils; D018408:Patch-Clamp Techniques; D014867:Water / Q000737:chemistry*
D005782:Gels; D008051:Lipid Bilayers; D008565:Membrane Proteins; D009821:Oils; D014867:Water