J. Lipid Res.  Neurobiology of Lipids (ISSN1683-5506)
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A more recent version of this article appeared on May 1, 2005

Papers In Press, published online ahead of print February 16, 2005
J. Lipid Res., doi:10.1194/jlr.D400041-JLR200
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Submitted on December 27, 2004
Revised on February 7, 2005
Accepted on February 7, 2005

A simplified method for the preparation of detergent-free lipid rafts

Jennifer L. Macdonald and Linda J. Pike

Department of Biochemistry and Molecular Biophysics, Washington University, St. Louis, MO 63110

Corresponding Author: pike{at}biochem.wustl.edu

Lipid rafts are small plasma membrane domains that contain high levels of cholesterol and sphingolipids. Traditional methods for the biochemical isolation of lipid rafts involve the extraction of cells with non-ionic detergents followed by the separation of a low density, detergent-resistant membrane fraction on density gradients. Because of concerns regarding the possible introduction of artifacts through the use of detergents, it is important to develop procedures for the isolation of lipid rafts that do not involve detergent extraction. We report here a simplified method for the purification of detergent-free lipid rafts that requires only one short density gradient centrifugation but yields a membrane fraction that is highly enriched in cholesterol and protein markers of lipid rafts with no contamination from non-raft plasma membrane or intracellular membranes.


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