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Isolation and Analysis of Detergent-Resistant Membrane Fractions

  • Massimo Aureli
  • Sara Grassi
  • Sandro Sonnino
  • Alessandro PrinettiEmail author
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1376)

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.

Key words

Detergent-resistant membrane Lipid raft Liquid-ordered phase Membrane domain Microdomain Sphingolipid 

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  • Massimo Aureli
    • 1
  • Sara Grassi
    • 1
  • Sandro Sonnino
    • 1
  • Alessandro Prinetti
    • 1
    Email author
  1. 1.Department of Medical Biotechnology and Translational MedicineUniversity of MilanSegrateItaly

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