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Kinetic Silver Staining of Proteins

  • Douglas D. Root
  • Kuan Wang
Protocol
  • 164 Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Silver staining methods have long been know to provide highly sensitive detection of proteins and nucleic acids following electrophoresis in agarose and polyacrylamide gels (1,2). Silver staining technologies can be extended to other media such as blots, thin-layer chromatography (TLC), and microtiter plates. The quantification of proteins adsorbed to microtiter plate wells provides quantitative information for enzyme-linked immunosorbent assay (ELISA) and protein interaction assays. One nonradioactive procedure, copper iodide staining, is described in  Chapter 51 (3). The kinetic silver staining method for measuring the amount of adsorbed protein in a microtiter plate has been developed recently. The microtiter plate assay has a sensitivity similar to copper iodide staining (5-150 ng/well) but higher precision (<5%; 4). When quantification is based on the time required for staining to reach a fixed optical density, very little protein-to-protein variation is observed, so a standard protein for calibration can be selected (such as bovine albumin) that is free of interfering substances to which the assay is sensitive (4). Furthermore, this kinetic silver staining assay is found to be most sensitive for the detection of proteins on cellulose such as is commonly used for TLC (see  Chapter 51 for a comparison with other solid-phase stains).

Keywords

Microtiter Plate Silver Staining Adsorbed Protein Standard Protein Microtiter Plate Reader 
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.
    Somerville, L. L. and Wang, K. (1981) The ultrasensitive silver &quote:protein&quote: stain also detects nanograms of nucleic acids. Biochem. Biophys. Res. Commun. 102, 53–58.PubMedCrossRefGoogle Scholar
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    Gottlieb, M. and Chavko, M. (1987) Silver staining of native and denatured eucaryotic DNA in agarose gels. Analyt. Biochem. 165, 33–37.PubMedCrossRefGoogle Scholar
  3. 3.
    Root, D. D. and Reisler, E. (1990) Copper iodide staining and determination of proteins adsorbed to microtiter plates. Analyt. Biochem. 186, 69–73.PubMedCrossRefGoogle Scholar
  4. 4.
    Root, D. D. and Wang, K. (1993) Kinetic silver staining and calibration of proteins adsorbed to microtiter plates. Analyt. Biochem. 209, 354–359.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Douglas D. Root
    • 1
  • Kuan Wang
    • 2
  1. 1.Chester College of Higher EducationUK
  2. 2.Department of Biological SciencesUniversity of North TexasDenton

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