title: Electroformation of giant unilamellar vesicles from native membranes and organic lipid mixtures for the study of lipid domains under physiological ionic-strength conditions.
abstract: Giant unilamellar vesicles (GUVs) constitute a cell-sized model membrane system that allows direct visualization of particular membrane-related phenomena, such as domain formation, at the level of single vesicles using fluorescence microscopy-related techniques. Currently available protocols for the preparation of GUVs work only at very low salt concentrations, thus precluding experimentation under physiological conditions. In addition, the GUVs thus obtained lack membrane compositional asymmetry. Here we show how to prepare GUVs using a new protocol based on the electroformation method either from native membranes or organic lipid mixtures at physiological ionic strength. Additionally, we describe methods to test whether membrane proteins and glycosphingolipids preserve their natural orientation after electroformation of GUVs composed of native membranes.
authors.affiliation: Unidad de Biofisica, CSIC-UPV/EHU, Leioa, Spain.
authors.forename: L-Ruth
authors.initials: LR
authors.lastname: Montes
authors.forename: Hasna
authors.initials: H
authors.lastname: Ahyayauch
authors.forename: Maitane
authors.initials: M
authors.lastname: Ibarguren
authors.forename: Jesus
authors.initials: J
authors.lastname: Sot
authors.forename: Alicia
authors.initials: A
authors.lastname: Alonso
authors.forename: Luis A
authors.initials: LA
authors.lastname: Bagatolli
authors.forename: Felix M
authors.initials: FM
authors.lastname: Goñi
mesh_terms: D002462:Cell Membrane / Q000737:chemistry* / Q000648:ultrastructure; D011211:Electric Power Supplies; D004910:Erythrocyte Membrane / Q000737:chemistry / Q000648:ultrastructure; D006801:Humans; D008055:Lipids / Q000737:chemistry; D008563:Membrane Lipids / Q000032:analysis* / Q000302:isolation & purification; D021962:Membrane Microdomains / Q000737:chemistry; D009994:Osmolar Concentration; D053835:Unilamellar Liposomes / Q000737:chemistry*
chemical_list: D008055:Lipids; D008563:Membrane Lipids; D053835:Unilamellar Liposomes