Constructing droplet interface bilayers from the contact of aqueous droplets in oil
| Autor: | Linda C. M. Gross, Oliver Kieran Castell, Sebastian Leptihn, Mark I. Wallace, Matthew A. Holden, David P. Marshall, James R. Thompson, Bríd Cronin, En-Hsin Lee |
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| Jazyk: | angličtina |
| Rok vydání: | 2016 |
| Předmět: |
Silver
Bacterial Toxins Lipid Bilayers 02 engineering and technology 010402 general chemistry 01 natural sciences Ion Channels General Biochemistry Genetics and Molecular Biology Hemolysin Proteins Mice Alkanes Monolayer Animals Lipid bilayer phase behavior Lipid bilayer Ion channel Antigens Bacterial Aqueous solution Chemistry Bilayer Silver Compounds Water Lipid bilayer fusion Membranes Artificial 021001 nanoscience & nanotechnology Nanostructures 0104 chemical sciences Membrane Chemical engineering 0210 nano-technology Oils |
| Popis: | We describe a protocol for forming an artificial lipid bilayer by contacting nanoliter aqueous droplets in an oil solution in the presence of phospholipids. A lipid monolayer forms at each oil-water interface, and when two such monolayers touch, a bilayer is created. Droplet interface bilayers (DIBs) are a simple way to generate stable bilayers suitable for single-channel electrophysiology and optical imaging from a wide variety of preparations, ranging from purified proteins to reconstituted eukaryotic cell membrane fragments. Examples include purified proteins from the α-hemolysin pore from Staphylococcus aureus, the anthrax toxin pore and the 1.2-MDa mouse mechanosensitive channel MmPiezo1. Ion channels and ionotropic receptors can also be reconstituted from membrane fragments without further purification. We describe two approaches for forming DIBs. In one approach, a lipid bilayer is created between two aqueous droplets submerged in oil. In the other approach, a membrane is formed between an aqueous droplet and an agarose hydrogel, which allows imaging in addition to electrical recordings. The protocol takes |
| Databáze: | OpenAIRE |
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