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J Neurochem. 2011 Mar;116(5):671-7. doi: 10.1111/j.1471-4159.2010.07076.x. Epub 2011 Jan 12.

Isolation of nano-meso scale detergent resistant membrane that has properties expected of lipid 'rafts'.

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  • 1Wolfson Centre for Age-related Disease, School of Biomedical Sciences, King's College London, London, UK. roger.morris@kcl.ac.uk

Abstract

This review assesses problems that confound attempts to isolate 'raft' domains from cell membranes, focusing in particular upon the isolation of detergent resistant membrane (DRM). Despite its widespread use, this technique is rightly viewed with skepticism by many membrane biochemists and biophysics for reasons that include the inability to isolate DRMs at 37°C, the temperature at which their lipids are supposed to be ordered and so exclude detergents. If solubilization is done in an ionic buffer that preserves the lamellar phase of the metastable inner leaflet lipids, DRMs can readily be isolated at 37°C, and these have many properties expected of lipid rafts. However, to date these DRMs have remained somewhat larger than current concepts of rafts. We describe an adaptation of this method that purifies nano-meso scale DRMs, and could be a significant step towards purifying the membrane of individual 'rafts'.

© 2011 The Authors. Journal of Neurochemistry © 2011 International Society for Neurochemistry.

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