Advertisement

Purification of Microgram Quantities of Proteins by Polyacrylamide Gel Electrophoresis

  • Michael W. Hunkapiller
  • Eva Lujan
Protocol
Part of the Biological Methods book series (BM)

Abstract

Polyacrylamide gel electrophoresis (PAGE) is one of the most widely used techniques for the analytical separation of proteins and peptides. PAGE separations, based upon intrinsic charge-to-mass ratio (1), apparent molecular weight in the presence of sodium dodecyl sulfate (SDS) (2), and isoelectric focusing (3), all possess remarkable resolving power using amounts of protein ranging from a few picograms to several milligrams. When two of these techniques, molecular sieving with SDS and isoelectric focusing with ampholytes, are combined in two-dimensional separations (4), complex mixtures can be resolved into as many as several thousand components in a single experiment.

Keywords

Sodium Dodecyl Sulfate Ammonium Bicarbonate Dialysis Membrane Fraction Collector Sodium Dodecyl Sulfate Solution 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Ornstein, L (1964) Ann. NY Acad. Sci. 121, 321–349.PubMedCrossRefGoogle Scholar
  2. 2.
    Laemmli, U. K. (1970) Nature (Lond) 227, 680–685.CrossRefGoogle Scholar
  3. 3.
    Catsimpoolas, N. (1970) Sep. Sci. 5, 523–545.CrossRefGoogle Scholar
  4. 4.
    O’Farrell, P. H. (1975) J. Biol. Chem. 250, 4007–4021.PubMedGoogle Scholar
  5. 5.
    Anderson, C. W., Baum, P. R., and Gesteland, R. F. (1973) J. Virol 12, 241–253.PubMedGoogle Scholar
  6. 6.
    Lazarides, E. (1976) J. Supramol. Struct. 5, 531–547.PubMedCrossRefGoogle Scholar
  7. 7.
    Hunkapiller, M. W., Lujan, E., Ostrander, F., and Hood, L. E. (1983) Methods Enzymol. 91, 227–236.PubMedCrossRefGoogle Scholar
  8. 8.
    Merril, C. R., Goldman, D., Sedman, S. A., and Ebert, M. H. (1981) Science 211, 1437–1438.PubMedCrossRefGoogle Scholar
  9. 9.
    Wessel, D. and Flugge, U. I. (1984) Anal. Biochem. 138, 141–143.PubMedCrossRefGoogle Scholar

Copyright information

© The Humana Press Inc. 1986

Authors and Affiliations

  • Michael W. Hunkapiller
    • 1
  • Eva Lujan
    • 2
  1. 1.Applied BiosystemsFoster City
  2. 2.California Institute of TechnologyPasadena

Personalised recommendations