title: Isolation and Analysis of Detergent-Resistant Membrane Fractions.
abstract: The hypothesis that the Golgi apparatus is capable of sorting proteins and sending them to the plasma membrane through "lipid rafts," membrane lipid domains highly enriched in glycosphingolipids, sphingomyelin, ceramide, and cholesterol, was formulated by van Meer and Simons in 1988 and came to a turning point when it was suggested that lipid rafts could be isolated thanks to their resistance to solubilization by some detergents, namely Triton X-100. An incredible number of papers have described the composition and properties of detergent-resistant membrane fractions. However, the use of this method has also raised the fiercest criticisms. In this chapter, we would like to discuss the most relevant methodological aspects related to the preparation of detergent-resistant membrane fractions, and to discuss the importance of discriminating between what is present on a cell membrane and what we can prepare from cell membranes in a laboratory tube.
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Fratelli Cervi 93, 20090, Segrate, Milano, Italy.
authors.forename: Massimo
authors.initials: M
authors.lastname: Aureli
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Fratelli Cervi 93, 20090, Segrate, Milano, Italy.
authors.forename: Sara
authors.initials: S
authors.lastname: Grassi
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Fratelli Cervi 93, 20090, Segrate, Milano, Italy.
authors.forename: Sandro
authors.initials: S
authors.lastname: Sonnino
authors.affiliation: Department of Medical Biotechnology and Translational Medicine, University of Milan, Via Fratelli Cervi 93, 20090, Segrate, Milano, Italy. alessandro.prinetti@unimi.it.
authors.forename: Alessandro
authors.initials: A
authors.lastname: Prinetti
mesh_terms: D002458:Cell Fractionation / Q000379:methods*; D002462:Cell Membrane / Q000737:chemistry* / Q000187:drug effects*; D003902:Detergents / Q000494:pharmacology*; D021962:Membrane Microdomains / Q000737:chemistry; D008565:Membrane Proteins / Q000737:chemistry; D013696:Temperature
chemical_list: D003902:Detergents; D008565:Membrane Proteins
keywords: Detergent-resistant membrane; Lipid raft; Liquid-ordered phase; Membrane domain; Microdomain; Sphingolipid