Fractionation of Subcellular Organelles

John M. Graham1

1 JG Research Consultancy, Wirral
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Unit 3.1
DOI:  10.1002/0471143030.cb0301s69
Online Posting Date:  December, 2015
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Abstract

This unit provides both a theoretical and a practical background to all the techniques associated with the application of differential and density gradient centrifugation for the analysis of subcellular membranes. The density gradient information focuses on the use of the modern gradient solute iodixanol, chosen for its ease of use, versatility, and compatibility with biological particles. Its use in both pre‐formed discontinuous and continuous gradients and in self‐generated gradients is discussed. Considerable emphasis is given to selection of the appropriate centrifuge rotors and tubes and their influence on the methods used for creation, fractionation, and analysis of density gradients. Without proper consideration of these critical ancillary procedures, the resolving power of the gradient can be easily compromised. © 2015 by John Wiley & Sons, Inc.

Keywords: differential centrifugation; density gradient technology; instrumentation

     
 
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Table of Contents

  • Introduction
  • Basic Principles of Centrifugation
  • Differential Centrifugation
  • Density Gradient Centrifugation
  • Making Gradients
  • Harvesting Gradients
  • Gradient Analysis
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
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