Dialysis and Ultrafiltration

Louis Zumstein1

1 Sennes Drug Innovations, Houston, Texas
Publication Name:  Current Protocols in Molecular Biology
Unit Number:  Appendix 3C
DOI:  10.1002/0471142727.mba03cs41
Online Posting Date:  May, 2001
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Abstract

The first section of this unit describes dialysis of large‐ and small‐volume samples using cellulose membranes with pore sizes designed to exclude molecules above a selected molecular weight. A support protocol describes preparation of membranes for dialysis. The second section describes protocols for ultrafiltration, and the third section describes simple methods of concentrating solutions.

     
 
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Table of Contents

  • Dialysis
  • Basic Protocol 1: Large‐Volume Dialysis
  • Alternate Protocol 1: Small‐Volume Dialysis
  • Support Protocol 1: Selection and Preparation of Dialysis Membrane
  • Ultrafiltration
  • Basic Protocol 2: Ultrafiltration Using Vacuum Dialysis
  • Alternate Protocol 2: Ultrafiltration Using Centrifuge Concentrators
  • Alternate Protocol 3: Ultrafiltration Using Stirred‐Cell Apparatus
  • Alternate Protocol 4: Ultrafiltration Using Tangential‐Flow Systems
  • Concentration Using Solid‐Phase or Slurry Absorbents
  • Key References
  • Figures
     
 
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Materials

Basic Protocol 1: Large‐Volume Dialysis

  Materials
    For common stock solutions, see appendix 22; for suppliers, see appendix 44.
  • Dialysis membrane (see protocol 3)
  • Clamps (Spectrapor Closures, Spectrum, or equivalent)
  • Macromolecule‐containing sample to be dialyzed
  • Appropriate dialysis buffer

Alternate Protocol 1: Small‐Volume Dialysis

  Additional Materials
  • Cork borer

Support Protocol 1: Selection and Preparation of Dialysis Membrane

  Additional Materials
    For common stock solutions, see appendix 22; for suppliers, see appendix 44.
  • 10 mM sodium bicarbonate
  • 10 mM Na 2EDTA, pH 8.0
  • 20% to 50% (v/v) ethanol

Basic Protocol 2: Ultrafiltration Using Vacuum Dialysis

  Materials
    For common stock solutions, see appendix 22; for suppliers, see appendix 44.
  • Sample
  • Appropriate dialysis buffer
  • Vacuum flask
  • One‐hole cork stopper
  • Pasteur pipet
  • Dialysis membrane of desired MWCO
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Figures

  •   FigureFigure a0.3C.1 Vacuum dialysis apparatus (from Craig 1967).

Videos

Literature Cited

Key References
   Craig, L.C. 1967. Techniques for the study of peptides and proteins by dialysis and diffusion. Methods Enzymol. 11:870‐905.
  Describe the theory and practice of dialysis and diffusion.
   McPhie, P. 1971. Dialysis. Methods Enzymol. 22:23‐32.
   ACS (American Chemical Society). 1994. Biotech Buyers' Guide 1994. ACS. Washington, D.C.
   McGregor, W.C. 1986. Selection and use of ultrafiltration membranes. In Membrane Separations in Biotechnology (W.C. McGregor, ed.) pp. 1‐36. Marcel Dekker, New York.
   Pohl, T. 1990. Concentration of proteins and removal of solutes. Methods Enzymol. 182:68‐83.
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