Analytical Ultracentrifugation

Thomas M. Laue1

1 University of New Hampshire, Durham
Publication Name:  Current Protocols in Protein Science
Unit Number:  Unit 7.5
DOI:  10.1002/0471140864.ps0705s04
Online Posting Date:  May, 2001
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Abstract

Analytical ultracentrifugation is one of the most powerful, though as yet underexploited, techniques available to molecular biology and biochemistry. This overview describes applications for analytical ultracentrifugation along with important considerations relating to experimental design.

     
 
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Table of Contents

  • Planning an Experiment
  • Anticipated Results
  • Time Considerations
  • Help for the Novice
  • Tables
     
 
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Materials

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Figures

Videos

Literature Cited

Literature Cited
   Attri, A.K. and Minton, A.P. 1986. Technique and apparatus for automated fractionation of the contents of small centrifuge tubes: Application to analytical ultracentrifugation. Anal. Biochem. 152:319‐328.
   Cohen, R. and Mire, M. 1971. Analytical‐band centrifugation of an active enzyme‐substrate complex. 1. Principle and practice of the centrifugation. Eur. J. Biochem. 23:267‐275.
   Harding, S.E., Rowe, A.J. and Horton, J.C. 1992. Analytical Ultracentrifugation in Biochemistry and Polymer Science. Royal Society of Chemistry, Cambridge.
   Laue, T.M., Shah, B., Ridgeway, T.M., and Pelletier, S.L. 1992. Computer‐aided interpretation of sedimentation data for proteins. In Analytical Ultracentrifugation in Biochemistry and Polymer Science (S.E. Harding, A.J. Rowe, and J.C. Horton, eds.) pp. 90‐125. Royal Society of Chemistry, Cambridge.
   Laue, T.M. 1992. Short Column Sedimentation Equilibrium Analysis for Rapid Characterization of Macromolecules in Solution. Technical Information DS‐835. Beckman Instruments, Palo Alto, Calif.
   Laue, T.M. 1995. Sedimentation equilibrium as a thermodynamic tool. Methods Enzymol. 259:427‐452.
   Mächtle, W. 1988. Coupling particle size technique. A new ultracentrifuge technique for determination of the particle size distribution of extremely broad distributed dispersions. Die Angew. Makromol. Chem. 162:35‐52.
   McRorie, D.K. and Voelker, P.J. 1993. Self Associating Systems in the Analytical Ultracentrifuge. Beckman Instruments, Fullerton, Calif.
   Philo, J.S. 1994. Measuring sedimentation, diffusion and molecular weights of small molecules by direct fitting of sedimentation velocity concentration profiles. In Modern Analytical Ultracentrifugation: Acquisition and Interpretation of Data for Biological and Synthetic Polymer Systems (T.M. Schuster and T.M. Laue, eds.) pp. 156‐170. Birkhauser, Boston.
   Ralston, G. 1993. Introduction to Analytical Ultracentrifugation. Beckman Instruments, Fullerton, Calif.
   Richards, E.G. and Schachman, H.K. 1957. A differential ultracentrifuge technique for measuring small changes in sedimentation coefficient. J. Am. Chem. 79:5324‐5325.
   Schachman, H.K. 1959. Ultracentrifugation in Biochemistry. Academic Press, New York.
   Schuster, T.M. and Laue, T.M. 1994. Modern Analytical Ultracentrifugation: Acquisition and Interpretation of Data for Biological and Synthetic Polymer Systems. Birkhauser, Boston.
   Stafford, W.F. III. 1992. Boundary analysis in sedimentation transport experiments: A procedure for obtaining sedimentation coefficient distributions using the time derivative of the concentration profile. Anal. Biochem. 161:70‐79.
   Stafford, W.F. III. 1994. Sedimentation boundary analysis of interacting systems. In Modern Analytical Ultracentrifugation: Acquisition and Interpretation of Data for Biological and Synthetic Polymer Systems (T.M. Schuster and T.M. Laue, eds.) pp. 119‐137. Birkhauser, Boston.
   Svedberg, T. and Pederson, K.O. 1940. The Ultracentrifuge. Clarendon Press, Oxford.
   Van Holde, K.E. and Weischet, W.O. 1978. Boundary analysis of sedimentation‐velocity experiments with monodisperse and paucidisperse solutes. Biopolymers 17:1387‐1403.
   Yphantis, D.A. 1960. Rapid determination of molecular weights of peptides and proteins. Ann. N.Y. Acad. Sci. 88:586‐601.
   Yphantis, D.A. 1964. Equilibrium ultracentrifugation of dilute solutions. Biochemistry 3:297‐317.
Key References
   Harding et al., 1992. See above.
  Excellent book containing descriptions of many of the modern methods of analytical ultracentrifugation.
   Schachman, 1959. See above.
  Classic work describing many useful methods still used today.
   Schuster and Laue, 1994. See above.
  Up‐to‐date collection of methods for analytical ultracentrifugation.
   Svedberg and Pederson, 1940. See above.
  A classic work that is still relevant in that it provides some of the clearest descriptions available of analytical centrifugation.
Internet Resources
  www.bbri.harvard.edu/rasmb/rasmb.html
  Web site for most recent programs and discussion group on analytical ultracentrifugation.
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