Advertisement

Silver Staining of 2-D Gels

  • Julia Poland
  • Thierry Rabilloud
  • Pranav Sinha
Protocol
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Proteomics, in its commonly used meaning, refers to the detection of differentially expressed proteins between control and experimental samples. The basic tools include two-dimensional gel electrophoresis (2-DE) for protein fractionation followed by mass spectrometry (MS) of excised protein spots for protein identification. After separation, proteins need to be detected on the gel, meaning the spot pattern has to be distinguished from the surrounding background. This is generally performed directly in the polyacrylamide gel by binding a dye molecule to the proteins.

Keywords

Silver Staining Silver Nitrate Stop Solution Spot Pattern Silver Sulfide 
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.
    Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal. Chem. 68, 850–858.PubMedCrossRefGoogle Scholar
  2. 2.
    Yan, J. X., Wait, R., Berkelman, T., et al. (2000) A modified silver staining protocol for visualization of proteins compatible with matrix-assisted laser desorption/ionization and electrospray ionization-mass spectrometry. Electrophoresis 21, 3666–3672.PubMedCrossRefGoogle Scholar
  3. 3.
    Sinha, P., Poland, J., Schnolzer, M., and Rabilloud, T. (2001) A new silver staining apparatus and procedure for matrix-assisted laser desorption/ionization-time of flight analysis of proteins after two-dimensional electrophoresis. Proteomics 1, 835–840.PubMedCrossRefGoogle Scholar
  4. 4.
    Rabilloud, T., Vuillard, L., Gilly, C., and Lawrence, J. J. (1994) Silver-staining of proteins in polyacrylamide gels: a general overview. Cell. Mol. Biol. (Noisy-le-grand) 40, 57–75.Google Scholar
  5. 5.
    Rabilloud, T. (1990) Mechanisms of protein silver staining in polyacrylamide gels: a 10-year synthesis. Electrophoresis 11, 785–794.PubMedCrossRefGoogle Scholar
  6. 6.
    Richert, S., Luche, S., Chevallet, M., Van Dorsselaer, A., Leize-Wagner, E., and Rabilloud, T. (2004) About the mechanism of interference of silver staining with peptide mass spectrometry. Proteomics 4, 909–916.PubMedCrossRefGoogle Scholar
  7. 7.
    Heukeshoven, J. and Dernick, R. (1985) Simplified method for silver staining of proteins in polyacrylamide gels and the mechanism of silver staining. Electrophoresis 6, 103–112.CrossRefGoogle Scholar
  8. 8.
    Klose, J. and Kobalz, U. (1995) Two-dimensional electrophoresis of proteins: an updated protocol and implications for a functional analysis of the genome. Electrophoresis 16, 1034–1059.PubMedCrossRefGoogle Scholar
  9. 9.
    Gharahdaghi, F., Weinberg, C. R., Meagher, D. A., Imai, B. S., and Mische, S. M. (1999) Mass spectrometric identification of proteins from silver-stained polyacrylamide gel: a method for the removal of silver ions to enhance sensitivity. Electrophoresis 20, 601–605.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2005

Authors and Affiliations

  • Julia Poland
    • 1
  • Thierry Rabilloud
    • 2
  • Pranav Sinha
    • 1
  1. 1.Institut fur Medizinische und Chemische LabordiagnostikLandeskrankenhaus KlagenfurtKlagenfurtAustria
  2. 2.DBMS/BECPCEA-GrenobleFrance

Personalised recommendations