Dialysis and Concentration of Protein Solutions

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

1 Lancaster University, Lancaster, 2 National Cancer Institute, Bethesda, Maryland, 3 Introgen Therapeutics, Inc., Houston, Texas
Publication Name:  Current Protocols in Immunology
Unit Number:  Appendix 3H
DOI:  10.1002/0471142735.ima03hs21
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. This appendix describes dialysis of large‐ and small‐volume samples using cellulose membranes with pore sizes designed to exclude molecules above a selected molecular weight. A 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
  • Key References
  • 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

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