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TheScientificWorldJOURNAL
Volume 2, Pages 1444-1448
http://dx.doi.org/10.1100/tsw.2002.839
Peer-Reviewed Protocol

OptiPrepTM Density Gradient Solutions for Nonmammalian Organelles

School of Biomolecular Sciences, Liverpool John Moores University, UK

Received 7 March 2002; Accepted 15 May 2002

Copyright © 2002 John M. Graham.

Abstract

Any density gradient for the isolation of nonmammalian organelles should ideally only expose the sedimenting biological particles to an increasing concentration of the gradient solute. Thus they will experience only an increasing density and viscosity, other parameters such as osmolality, pH, ionic strength and the concentration of important additives (such as EDTA and DTT) should remain as close to constant as possible. This Protocol Article describes the strategies for the dilution of OptiPrep™ in order to prepare such solutions for organelles and membranes from nonmammalian sources such as yeast.