Abstract
Silver staining is used to detect proteins after electrophoretic separation on polyacrylamide gels. It combines excellent sensitivity (in the low nanogram range) with the use of very simple and cheap equipment and chemicals. It is compatible with downstream processing, such as mass spectrometry analysis after protein digestion. 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 d after the end of the electrophoretic separation. Once completed, the stain is stable for several weeks.
This is a preview of subscription content, access via your institution
Access options
Subscribe to this journal
Receive 12 print issues and online access
$259.00 per year
only $21.58 per issue
Buy this article
- Purchase on Springer Link
- Instant access to full article PDF
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Rabilloud, T. Mechanisms of protein silver staining in polyacrylamide gels: a 10-year synthesis. Electrophoresis 11, 785–794 (1990).
Merril, C.R. et al. Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211, 1437–1438 (1981).
Schagger, H. & von Jagow, G. Tricine-sodium dodecyl sulfate-polyacrylamide gel electrophoresis for the separation of proteins in the range from 1 to 100 kDa. Anal. Biochem. 166, 368–379 (1987).
Wiltfang, J. et al. A new multiphasic buffer system for sodium dodecyl sulfate-polyacrylamide gel electrophoresis of proteins and peptides with molecular masses 100,000–1000, and their detection with picomolar sensitivity. Electrophoresis 12, 352–366 (1991).
Tastet, C. et al. A versatile electrophoresis system for the analysis of high- and low-molecular weight proteins. Electrophoresis 24, 1787–1794 (2003).
Hochstrasser, D.F. & Merril, C.R. 'Catalysts' for polyacrylamide gel polymerization and detection of proteins by silver staining. Appl. Theor. Electrophor. 1, 35–40 (1988).
Eschenbruch, M. & Bürk, R.R. Experimentally improved reliability of ultrasensitive silver staining of protein in polyacrylamide gels. Anal. Biochem. 125, 96–99 (1982).
Chevallet, M. et al. Improved mass spectrometry compatibility is afforded by ammoniacal silver staining. Proteomics 6, 2350–2354 (2006).
Blum, H. et al. Improved silver staining of plant proteins, RNA and DNA in polyacrylamide gels. Electrophoresis 8, 93–99 (1987).
Rabilloud, T. A comparison between low background silver diammine and silver nitrate protein stains. Electrophoresis 13, 429–439 (1992).
Richert, S. et al. About the mechanism of interference of silver staining with peptide mass spectrometry. Proteomics 4, 909–916 (2004).
Sinha, P. et al. A new silver staining apparatus for MALDI/TOF analysis of proteins after two-dimensional electrophoresis. Proteomics 1, 835–840 (2001).
Mold, D.E. et al. Silver staining of histones in Triton-acid-urea gels. Anal. Biochem. 135, 44–47 (1983).
Rabilloud, T. et al. Two-dimensional electrophoresis of human placental mitochondria and protein ientification by mass spectrometry: Toward a human mitochondrial proteome. Electrophoresis 19, 1006–1014 (1998).
Gharahdaghi, F. et al. 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 (1999).
Neuhoff, V. et al. Improved staining of proteins in polyacrylamide gels including isoelectric focusing gels with clear background at nanogram sensitivity using Coomassie Brilliant Blue G-250 and R-250. Electrophoresis 9, 255–262 (1988).
Author information
Authors and Affiliations
Corresponding author
Ethics declarations
Competing interests
The authors declare no competing financial interests.
Rights and permissions
About this article
Cite this article
Chevallet, M., Luche, S. & Rabilloud, T. Silver staining of proteins in polyacrylamide gels. Nat Protoc 1, 1852–1858 (2006). https://doi.org/10.1038/nprot.2006.288
Published:
Issue Date:
DOI: https://doi.org/10.1038/nprot.2006.288
This article is cited by
-
Identification of the protein coding capability of coronavirus defective viral genomes by mass spectrometry
Virology Journal (2023)
-
Immunoproteomic analysis of the serum IgG response to cell wall-associated proteins of Staphylococcus aureus strains belonging to CC97 and CC151
Veterinary Research (2023)
-
Assessment of genetic diversity among sago palms (Metroxylon sagu Rottb.) in Bengkulu, Indonesia using simple sequence repeats
Genetic Resources and Crop Evolution (2023)
-
The effect of temperature on the activity and stability of the thermostable enzyme caffeine dehydrogenase from Pichia manshurica CD1
Biologia (2023)
-
Peptide Models of the Cytoplasmic Tail of Influenza A/H1N1 Virus Hemagglutinin Expand Understanding its pH-Dependent Modes of Interaction with Matrix Protein M1
The Protein Journal (2023)
Comments
By submitting a comment you agree to abide by our Terms and Community Guidelines. If you find something abusive or that does not comply with our terms or guidelines please flag it as inappropriate.
