Skip to main content
SpringerLink
Log in
Book cover

Two-Dimensional Electrophoresis Protocols pp 339–350Cite as

Silver Staining of Proteins in 2DE Gels

  • Protocol
  • First Online:

Part of the book series: Methods in Molecular Biology ((MIMB,volume 519))

Summary

Silver staining detects proteins after electrophoretic separation on polyacrylamide gels. Its main positive features are its excellent sensitivity (in the low nanogram range) and the use of very simple and cheap equipment and chemicals. The sequential phases of silver staining are protein fixation, then sensitization, then silver impregnation, and finally image development. Several variants of silver staining are described here, which can be completed in a time range from 2 h to 1 day after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.

This is a preview of subscription content, log in via an institution.

Protocol
USD   49.95
Price excludes VAT (USA)
  • Available as PDF
  • Read on any device
  • Instant download
  • Own it forever
eBook
USD   129.00
Price excludes VAT (USA)
  • Available as EPUB and PDF
  • Read on any device
  • Instant download
  • Own it forever
Softcover Book
USD   169.00
Price excludes VAT (USA)
  • Compact, lightweight edition
  • Dispatched in 3 to 5 business days
  • Free shipping worldwide - see info

Tax calculation will be finalised at checkout

Purchases are for personal use only

Learn about institutional subscriptions

Springer Nature is developing a new tool to find and evaluate Protocols. Learn more

References

  1. Switzer, R.C., Merril, C.R., and Shifrin, S. (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal. Biochem. 98, 231–237

    Article  PubMed  CAS  Google Scholar 

  2. Rabilloud, T. (1990) Mechanisms of protein silver staining in polyacrylamide gels: a ten years synthesis. Electrophoresis 11, 785–794

    Article  PubMed  CAS  Google Scholar 

  3. Rabilloud, T., Vuillard, L., Gilly, C., and Lawrence, J.J. (1994) Silver staining of proteins in polyacrylamide gels: a general overview. Cell. Mol. Biol. 40, 57–75

    PubMed  CAS  Google Scholar 

  4. Blum, H., Beier, H., and Gross, H.J. (1987) Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8, 93–99

    Article  CAS  Google Scholar 

  5. Hochstrasser, D.F., and Merril, C.R. (1988) Catalysts for polyacrylamide gel polymerization and detection of proteins by silver staining. Appl. Theor. Electrophor. 1, 35–40

    PubMed  CAS  Google Scholar 

  6. Wiederkehr, F., Ogilvie, A., and Vonderschmitt, D. (1985) Two-dimensional gel electrophoresis of cerebrospinal fluid proteins from patients with various neurological diseases. Clin. Chem. 31, 1537–1542

    PubMed  CAS  Google Scholar 

  7. 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

    Article  PubMed  CAS  Google Scholar 

  8. Metz, B., Kersten, G.F., Baart, G.J., de Jong, A., Meiring, H., ten Hove, J., et al. (2006) Identification of formaldehyde-induced modifications in proteins: reactions with insulin. Bioconjug. Chem. 17, 815–822

    Article  PubMed  CAS  Google Scholar 

  9. Oses-Prieto, J.A., Zhang, X., and Burlingame, A.L. (2007) Formation of {epsilon}-formyllysine on silver-stained proteins: Implications for assignment of isobaric dimethylation sites by tandem mass spectrometry. Mol. Cell. Proteomics 6, 181–192

    PubMed  CAS  Google Scholar 

  10. Chevallet, M., Diemer, H., Luche, S., van Dorsselaer, A., Rabilloud, T., and Leize-Wagner, E. (2006) Improved mass spectrometry compatibility is afforded by ammoniacal silver staining. Proteomics 6, 2350–2354

    Article  PubMed  CAS  Google Scholar 

  11. Sinha, P., Poland, J.,Schnolzer, M., andRabilloud, T. (2001) A new silver staining apparatus for MALDI/TOF analysis of proteins after two-dimensional electrophoresis Proteomics 1, 835–840

    Article  PubMed  CAS  Google Scholar 

  12. Eschenbruch, M., and Bürk, R.R. (1982) Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal. Biochem. 125, 96–99

    Article  PubMed  CAS  Google Scholar 

  13. Hochstrasser, D.F., Patchornik, A., and Merril, C.R. (1988) Development of polyacrylamide gels that improve the separation of proteins and their detection by silver staining. Anal. Biochem. 173, 412–423

    Article  PubMed  CAS  Google Scholar 

  14. Rabilloud, T. (1992) A comparison between low background silver diammine and silver nitrate protein stains. Electrophoresis 13, 429–439

    Article  PubMed  CAS  Google Scholar 

  15. Ochs, D.C., McConkey, E.H., and Sammons, D.W. (1981) Silver staining for proteins in polyacrylamide gels: A comparison of six methods. Electrophoresis 2, 304–307

    Article  CAS  Google Scholar 

  16. Shevchenko, A., Wilm, M., Vorm, O., and Mann, M. (1996) Mass spectrometric sequencing of proteins silver-stained polyacrylamide gels. Anal Chem. 68, 850–858

    Article  PubMed  CAS  Google Scholar 

Download references

Author information

Authors and Affiliations

  1. CEA-DSV-iRTSV/LBBSI and UMR CNRS 5092, CEA Grenoble, 17 rue des martyrs, F-38054, Grenoble Cedex 9, France

    Cécile Lelong, Mireille Chevallet, Sylvie Luche & Thierry Rabilloud

Authors
  1. Cécile Lelong
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mireille Chevallet
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Sylvie Luche
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thierry Rabilloud
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Thierry Rabilloud .

Editor information

Editors and Affiliations

    Rights and permissions

    Reprints and permissions

    Copyright information

    © 2009 Humana Press, a part of Springer Science+Business Media, LLC

    About this protocol

    Cite this protocol

    Lelong, C., Chevallet, M., Luche, S., Rabilloud, T. (2009). Silver Staining of Proteins in 2DE Gels. In: Tyther, R., Sheehan, D. (eds) Two-Dimensional Electrophoresis Protocols. Methods in Molecular Biology, vol 519. Humana Press. https://doi.org/10.1007/978-1-59745-281-6_21

    Download citation

    • DOI: https://doi.org/10.1007/978-1-59745-281-6_21

    • Published:

    • Publisher Name: Humana Press

    • Print ISBN: 978-1-58829-937-6

    • Online ISBN: 978-1-59745-281-6

    • eBook Packages: Springer Protocols

    Publish with us

    Policies and ethics

    Search

    Navigation