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Volume 2, Pages 1662-1666
Peer-Reviewed Protocol

Purification of Lipid Rafts from Cultured Cells

School of Biomolecular Sciences, Liverpool John Moores University, UK

Received 7 March 2002; Revised 14 May 2002; Accepted 15 May 2002

Copyright © 2002 John Graham.


Lipid-rich lipid rafts are microdomains of the plasma membrane that are resistant to low concentrations of nonionic detergent. This forms the basis for their isolation. Either a microsomal fraction or a postnuclear supernatant are loaded beneath a discontinuous iodixanol gradient. If all the solutions contain 0.5–1.0% Triton X-100, the intact lipid rafts float to the top of the gradient while all of the other detergent-solubilized membranes remain at the bottom.