Dialysis and Concentration of Protein Solutions

Sarah. M. Andrew1, Julie. A. Titus2, Louis Zumstein3

1 Chester College, Chester, 2 National Cancer Institute, Bethesda, Maryland, 3 Introgen Therapeutics, Inc., Houston, Texas
Publication Name:  Current Protocols in Cell Biology
Unit Number:  Appendix 3C
DOI:  10.1002/0471143030.cba03cs04
Online Posting Date:  May, 2001
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Abstract

Conventional dialysis separates small molecules from large molecules by allowing diffusion of only the small molecules through selectively permeable membranes. Dialysis is usually used to change the salt (small‐molecule) composition of a macromolecule‐containing solution. 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 and discusses issues related to the selection of membranes including commercial kits.

     
 
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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
  • Basic Protocol 2: Concentration
  • Figures
     
 
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Materials

Basic Protocol 1: Large‐Volume Dialysis

  Materials
  • Dialysis membrane (see protocol 3)
  • Clamps (Spectra/Por Closures, Spectrum, or equivalent)
  • Macromolecule‐containing sample to be dialyzed
  • Appropriate dialysis buffer

Alternate Protocol 1: Small‐Volume Dialysis

  • 0.5‐ml microcentrifuge tube
  • Pasteur pipet
  • Cork borer

Support Protocol 1: Selection and Preparation of Dialysis Membrane

  Materials
    Dialysis membrane
  • 10 mM sodium bicarbonate
  • 10 mM Na 2EDTA, pH 8.0
  • 20% to 50% (v/v) ethanol
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Figures

Videos

Literature Cited

Literature Cited
   Craig, L.C. 1967. Techniques for the study of peptides and proteins by dialysis and diffusion. Methods Enzymol. 11:870‐905.
Key References
   Craig. 1967. See above.
  Describes the theory and practice of dialysis and diffusion.
   McPhie, P. 1971. Dialysis. Methods Enzymol. 22:23‐32.
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