title: Isolating HeLa cell fractions enriched for clathrin-coated vesicles.
abstract: HeLa cell lines can be experimentally manipulated using drugs or gene-silencing techniques such as RNA interference. Fractions enriched for clathrin-coated vesicles (CCVs) can be isolated from these cell lines and used to study the effects of these manipulations on the composition of CCVs. This protocol, originally developed in the laboratory of Margaret Robinson (Cambridge, United Kingdom), describes the preparation of a HeLa cell fraction that is enriched for a mixed population of CCVs and is suitable for analysis by mass spectroscopy, western blotting, or electron microscopy.
authors.affiliation: Department of Proteomics and Signal Transduction, Max Plank Institute of Biochemistry, 82152 Martinsried, Germany;
authors.forename: Georg H H
authors.initials: GH
authors.lastname: Borner
authors.affiliation: Department of Cellular and Molecular Physiology, Institute of Translational Medicine, University of Liverpool, Liverpool L69 3BX, United Kingdom.
authors.forename: Andrew B
authors.initials: AB
authors.lastname: Fielding
mesh_terms: D015153:Blotting, Western / Q000379:methods; D002458:Cell Fractionation / Q000379:methods*; D022163:Clathrin-Coated Vesicles / Q000737:chemistry* / Q000648:ultrastructure*; D006367:HeLa Cells; D006801:Humans; D013058:Mass Spectrometry / Q000379:methods; D008854:Microscopy, Electron