Fractionation of subcellular membrane vesicles of epithelial and nonepithelial cells by optiPrep™ density gradient ultracentrifugation

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Density gradient ultracentrifugation (DGUC) is widely used for physical isolation (enrichment rather than purification) of subcellular membrane vesicles. It has been a valuable tool to study specific subcellular localization and dynamic trafficking of proteins. While sucrose has been the main component of density gradients, a few years ago synthetic OptiPrep (iodixanol) began being used for separation of organelles because of its iso-osmotic property. Here, we describe a detailed protocol for density gradient fractionation of various mammalian subcellular vesicles, including endoplasmic reticulum (ER), Golgi apparatus, endosomes, and lipid rafts, as well as apical and basolateral membranes of polarized epithelial cells.

Original languageEnglish (US)
Title of host publicationExocytosis and Endocytosis
PublisherHumana Press
Pages97-110
Number of pages14
ISBN (Print)9781588298652
DOIs
StatePublished - 2008

Publication series

NameMethods in Molecular Biology
Volume440
ISSN (Print)1064-3745

Keywords

  • Density gradient ultracentrifugation
  • Epithelia
  • Lipid rafts
  • OptiPrep
  • Organelles
  • Subcellular membrane vesicles

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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