Advertisement

SDS-Polyacrylamide Gel Electrophoresis of Peptides

  • Ralph C. Judd
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) has proven to be among the most useful tools yet developed in the area of molecular biology. The discontinuous buffer system, first described by Laemmli (1), has made it possible to separate, visualize, and compare readily the component parts of complex mixtures of molecules (e.g., tissues, cells). SDS-PAGE separation of proteins and peptides makes it possible to quantify the amount of a particular protein/peptide in a sample, obtain fairly reliable molecular mass information, and, by combining SDS-PAGE with immunoelectroblotting, evaluate the antigenicity of proteins and peptides. SDS-PAGE is both a powerful separation system and a reliable preparative purification technique (2 and see Chapter 11).

Keywords

Ammonium Persulfate Coomassie Brilliant Blue Back Plate Front Plate Discontinuous Buffer System 
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.
    Laemmli, U. K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–695.PubMedCrossRefGoogle Scholar
  2. 2.
    Judd, R. C. (1988) Purification of outer membrane proteins of the Gram negative bacterium Neisseria gonorrhoeae. Anal. Biochem. 173, 307–316.PubMedCrossRefGoogle Scholar
  3. 3.
    Dreyfuss, G., Adam, S. A., and Choi, Y. D. (1984) Physical change in cytoplasmic messenger ribonucleoproteins in cells treated with inhibitors of mRNA transcription. Mol. Cell. Biol. 4, 415–423.PubMedGoogle Scholar
  4. 4.
    Schagger, H. and von Jagow, G. (1987) Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166, 368–397.PubMedCrossRefGoogle Scholar
  5. 5.
    Judd, R. C. (1986) Evidence for N-terminal exposure of the PIA subclass of protein I of Neisseria gonorrhoeae. Infect. Immunol. 54, 408–414.Google Scholar
  6. 6.
    Moos, M., Jr. and Nguyen, N. Y. (1988) Reproducible high-yield sequencing of proteins electrophoretically separated and transferred to an inert support. J. Biol. Chem. 263, 6005–6008.PubMedGoogle Scholar
  7. 7.
    Stoll, V. S. and Blanchard, J. S. (1990) Buffers: principles and practice, in Methods in Enzymology, vol. 182, A Guide to Protein Purification (Deutscher, M. P., ed.), Academic, San Diego, CA, pp. 24–38.Google Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Ralph C. Judd
    • 1
  1. 1.Division of Biological SciencesUniversity of MontanaMissoula

Personalised recommendations