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TheScientificWorldJOURNAL
Volume 2 (2002), Pages 1435-1439
http://dx.doi.org/10.1100/tsw.2002.286
Peer-Reviewed Protocol

Fractionation of Golgi, Endoplasmic Reticulum, and Plasma Membrane from Cultured Cells in a Preformed Continuous Iodixanol Gradient

School of Biomolecular Sciences, Liverpool John Moores University, USA

Received 7 March 2002; Accepted 15 May 2002

Copyright © 2002 John¬†Graham.

Abstract

A continuous iodixanol gradient within the range 0-30% (w/v) iodixanol can resolve the major membrane compartments of the endoplasmic reticulum, Golgi membranes, and plasma membrane from a postnuclear supernatant prepared from a cultured cell homogenate. The precise density range of the gradient and the centrifugation conditions (100,000-200,000 g for 2-16 h) vary with the type of cell and the requirements of the separation. The strategy is widely used to study the processing of proteins within cells.