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Reconstitution of mammalian ion channels in droplet bilayers

Abstract

Ion channel studies are important for scientific characterization of cellular processes and for the purpose of drug discovery. The sessile droplet bilayer platform allows for simple, fast, inexpensive and high-yield formation of artificial lipid bilayers and reconstitution of ion channels. We validated this bilayer formation platform with mammalian ion channels like Chloride Intracellular Channel 1 (CLIC1) and Voltage-gated potassium channel 1.2 (Kv1.2). Ion channel synthesis using traditional bacterial expression systems is a complex and time consuming process and it limits the number of eukaryotic proteins that can be expressed therein. We explored the use of in vitro protein synthesis and plasma membrane fractionation of commercially available mammalian cells for expressing and reconstituting ion channel proteins in planar lipid bilayers.

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