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Principle and Method of Silver Staining of Proteins Separated by Sodium Dodecyl Sulfate–Polyacrylamide Gel Electrophoresis

  • Gaurav Kumar
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 1853)

Abstract

Silver staining is an excellent technique for detecting proteins which are separated by sodium dodecyl sulfate–polyacrylamide gel electrophoresis due to its efficiency of detecting proteins present in nanograms. The technique is based on the simple principle that selective reduction of silver into metallic silver occurs at the initiation site in the close proximity of protein molecules. The staining process sequentially consists of protein fixation, sensitization, washing, silver impregnation, and finally development of image. Depending upon the amount of silver incorporated into the protein bands, different color of gel is produced on silver staining. Though different protocols of silver staining exist, the method described here is easy, cheap, reliable, and very sensitive.

Key words

Silver staining Proteins Polyacrylamide gels Fixation Development SDS-PAGE electrophoresis 

References

  1. 1.
    Westermeier R (2006) Sensitive, quantitative, and fast modifications for Coomassie Blue staining of polyacrylamide gels. Proteomics 6(Suppl 2):61–64CrossRefGoogle Scholar
  2. 2.
    Zehr BD, Savin TJ, Hall RE (1989) A one-step, low background coomassie staining procedure for polyacrylamide gels. Anal Biochem 182(1):157–159CrossRefGoogle Scholar
  3. 3.
    Kerenyi L, Gallyas F (1972) A highly sensitive method for demonstrating proteins in electrophoretic, immunoelectrophoretic and immunodiffusion preparations. Clin Chim Acta 38(2):465–467CrossRefGoogle Scholar
  4. 4.
    Switzer RC III, Merril CR, Shifrin S (1979) A highly sensitive silver stain for detecting proteins and peptides in polyacrylamide gels. Anal Biochem 98(1):231–237CrossRefGoogle Scholar
  5. 5.
    Merril CR et al (1981) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variation in cerebrospinal fluid proteins. Science 211(4489):1437–1438CrossRefGoogle Scholar
  6. 6.
    Rabilloud T (1992) A comparison between low background silver diammine and silver nitrate protein stains. Electrophoresis 13(7):429–439CrossRefGoogle Scholar
  7. 7.
    Merril CR et al (1988) Coloration of silver-stained protein bands in polyacrylamide gels is caused by light scattering from silver grains of characteristic sizes. Proc Natl Acad Sci U S A 85(2):453–457CrossRefGoogle Scholar
  8. 8.
    Hempelmann E, Krafts K (2017) The mechanism of silver staining of proteins separated by SDS polyacrylamide gel electrophoresis. Biotech Histochem 92(2):79–85CrossRefGoogle Scholar
  9. 9.
    Hempelmann E et al (1987) Studies on glutathione reductase and methemoglobin from human erythrocytes parasitized with Plasmodium falciparum. Mol Biochem Parasitol 23(1):19–24CrossRefGoogle Scholar
  10. 10.
    Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72:248–254CrossRefGoogle Scholar
  11. 11.
    Hempelmann E, Kaminsky R (1986) Long term stability of colors after silver staining. Electrophoresis 7(10):481–481CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2018

Authors and Affiliations

  1. 1.Oklahoma Medical Research Foundation, University of OklahomaOklahoma CityUSA

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