HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) All Versions of this Article: D400041-JLR200v1 46/5/1061    most recent Alert me when this article is cited Alert me if a correction is posted Citation Map Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Macdonald, J. L. Articles by Pike, L. J. Search for Related Content PubMed PubMed Citation Articles by Macdonald, J. L. Articles by Pike, L. J. Social Bookmarking                      What's this?

Journal of Lipid Research, Vol. 46, 1061-1067, May 2005
Copyright © 2005 by American Society for Biochemistry and Molecular Biology

Methods

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

Jennifer L. Macdonald and Linda J. Pike¹

Department of Biochemistry and Molecular Biophysics, Washington University School of Medicine, 660 South Euclid Avenue, St. Louis, MO 63110

¹ To whom correspondence should be addressed. e-mail: pike{at}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 nonionic 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 nonraft plasma membrane or intracellular membranes.

Abbreviations: CHO, Chinese hamster ovary; EGF, epidermal growth factor; ER, endoplasmic reticulum; GPI, glycosylphosphatidylinositol

Supplementary key words caveolin • cholesterol • epidermal growth factor receptors • flotillin • Gq

CiteULike    Complore    Connotea    Del.icio.us    Digg    Reddit    Technorati    What's this?

This article has been cited by other articles:

				E. G. Hofman, M. O. Ruonala, A. N. Bader, D. van den Heuvel, J. Voortman, R. C. Roovers, A. J. Verkleij, H. C. Gerritsen, and P. M. P. van Bergen en Henegouwen EGF induces coalescence of different lipid rafts J. Cell Sci., August 1, 2008; 121(15): 2519 - 2528. [Abstract] [Full Text] [PDF]

				H. Zheng, J. Chu, Y. Qiu, H. H. Loh, and P.-Y. Law Agonist-selective signaling is determined by the receptor location within the membrane domains PNAS, July 8, 2008; 105(27): 9421 - 9426. [Abstract] [Full Text] [PDF]

				M. J. Polyak, H. Li, N. Shariat, and J. P. Deans CD20 Homo-oligomers Physically Associate with the B Cell Antigen Receptor: DISSOCIATION UPON RECEPTOR ENGAGEMENT AND RECRUITMENT OF PHOSPHOPROTEINS AND CALMODULIN-BINDING PROTEINS J. Biol. Chem., July 4, 2008; 283(27): 18545 - 18552. [Abstract] [Full Text] [PDF]

				A. G. Ayuyan and F. S. Cohen Raft Composition at Physiological Temperature and pH in the Absence of Detergents Biophys. J., April 1, 2008; 94(7): 2654 - 2666. [Abstract] [Full Text] [PDF]

				X. Zhang, F. Tan, Y. Zhang, and R. A. Skidgel Carboxypeptidase M and Kinin B1 Receptors Interact to Facilitate Efficient B1 Signaling from B2 Agonists J. Biol. Chem., March 21, 2008; 283(12): 7994 - 8004. [Abstract] [Full Text] [PDF]

				S. Kannan, A. Audet, H. Huang, L.-j. Chen, and M. Wu Cholesterol-Rich Membrane Rafts and Lyn Are Involved in Phagocytosis during Pseudomonas aeruginosa Infection J. Immunol., February 15, 2008; 180(4): 2396 - 2408. [Abstract] [Full Text] [PDF]

				D. P. Morris, B. Lei, Y.-X. Wu, G. A. Michelotti, and D. A. Schwinn The {alpha}1a-Adrenergic Receptor Occupies Membrane Rafts with Its G Protein Effectors but Internalizes via Clathrin-coated Pits J. Biol. Chem., February 1, 2008; 283(5): 2973 - 2985. [Abstract] [Full Text] [PDF]

				M. Imai, K. Kawasaki, and T. Odagiri Cytoplasmic Domain of Influenza B Virus BM2 Protein Plays Critical Roles in Production of Infectious Virus J. Virol., January 15, 2008; 82(2): 728 - 739. [Abstract] [Full Text] [PDF]

				V. Campana, A. Caputo, D. Sarnataro, S. Paladino, S. Tivodar, and C. Zurzolo Characterization of the Properties and Trafficking of an Anchorless Form of the Prion Protein J. Biol. Chem., August 3, 2007; 282(31): 22747 - 22756. [Abstract] [Full Text] [PDF]

				R. Latif, T. Ando, and T. F. Davies Lipid Rafts Are Triage Centers for Multimeric and Monomeric Thyrotropin Receptor Regulation Endocrinology, July 1, 2007; 148(7): 3164 - 3175. [Abstract] [Full Text] [PDF]

				S. Tikku, Y. Epshtein, H. Collins, A. J. Travis, G. H. Rothblat, and I. Levitan Relationship between Kir2.1/Kir2.3 activity and their distributions between cholesterol-rich and cholesterol-poor membrane domains Am J Physiol Cell Physiol, July 1, 2007; 293(1): C440 - C450. [Abstract] [Full Text] [PDF]

				K. B. Kannan, D. Barlos, and C. J. Hauser Free Cholesterol Alters Lipid Raft Structure and Function Regulating Neutrophil Ca2+ Entry and Respiratory Burst: Correlations with Calcium Channel Raft Trafficking J. Immunol., April 15, 2007; 178(8): 5253 - 5261. [Abstract] [Full Text] [PDF]

				C. Vijayasarathy, Y. Takada, Y. Zeng, R. A. Bush, and P. A. Sieving Retinoschisin Is a Peripheral Membrane Protein with Affinity for Anionic Phospholipids and Affected by Divalent Cations Invest. Ophthalmol. Vis. Sci., March 1, 2007; 48(3): 991 - 1000. [Abstract] [Full Text] [PDF]

				M. Koseki, K.-i. Hirano, D. Masuda, C. Ikegami, M. Tanaka, A. Ota, J. C. Sandoval, Y. Nakagawa-Toyama, S. B. Sato, T. Kobayashi, et al. Increased lipid rafts and accelerated lipopolysaccharide-induced tumor necrosis factor-{alpha} secretion in Abca1-deficient macrophages J. Lipid Res., February 1, 2007; 48(2): 299 - 306. [Abstract] [Full Text] [PDF]

				C. M. Mutch, R. Sanyal, T. L. Unruh, L. Grigoriou, M. Zhu, W. Zhang, and J. P. Deans Activation-induced endocytosis of the raft-associated transmembrane adaptor protein LAB/NTAL in B lymphocytes: evidence for a role in internalization of the B cell receptor Int. Immunol., January 1, 2007; 19(1): 19 - 30. [Abstract] [Full Text] [PDF]

				F. Stetzkowski-Marden, K. Gaus, M. Recouvreur, A. Cartaud, and J. Cartaud Agrin elicits membrane lipid condensation at sites of acetylcholine receptor clusters in C2C12 myotubes J. Lipid Res., October 1, 2006; 47(10): 2121 - 2133. [Abstract] [Full Text] [PDF]

				M. Garcia-Marcos, E. Perez-Andres, S. Tandel, U. Fontanils, A. Kumps, E. Kabre, A. Gomez-Munoz, A. Marino, J.-P. Dehaye, and S. Pochet Coupling of two pools of P2X7 receptors to distinct intracellular signaling pathways in rat submandibular gland J. Lipid Res., April 1, 2006; 47(4): 705 - 714. [Abstract] [Full Text] [PDF]

				A. Maguy, T. E. Hebert, and S. Nattel Involvement of lipid rafts and caveolae in cardiac ion channel function Cardiovasc Res, March 1, 2006; 69(4): 798 - 807. [Abstract] [Full Text] [PDF]

				L. J. Pike, X. Han, and R. W. Gross Epidermal Growth Factor Receptors Are Localized to Lipid Rafts That Contain a Balance of Inner and Outer Leaflet Lipids: A SHOTGUN LIPIDOMICS STUDY J. Biol. Chem., July 22, 2005; 280(29): 26796 - 26804. [Abstract] [Full Text] [PDF]

				K. Gaus, M. Rodriguez, K. R. Ruberu, I. Gelissen, T. M. Sloane, L. Kritharides, and W. Jessup Domain-specific lipid distribution in macrophage plasma membranes J. Lipid Res., July 1, 2005; 46(7): 1526 - 1538. [Abstract] [Full Text] [PDF]