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Kinetic Silver Staining of Proteins Adsorbed to Microtiter Plates

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

Abstract

The quantification of proteins adsorbed on microtiter plates is useful for quantitative ELISA and protein interaction assays. One nonradioactive procedure, copper iodide staining, has been described (1 and see Chapter 8). A kinetic silver staining method for measuring the amount of adsorbed protein in a microtiter plate has been developed. This assay has a sensitivity similar to copper iodide staining (5–150 ng/well), but much higher precision (<5%; 2). The kinetic silver staining assay uses the silver staining reagent developed originally by Gottlieb and Chavko for nucleic acids in agarose gels (3). Quantification is based on the time required for staining to reach a fixed optical density. This time-based assay has very little protein-to-protein variation, but is sensitive to interfering substances (2).

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.
    Root, D. D. and Reisier, E. (1990) Copper iodide staining and determination of proteins adsorbed to microtiter plates. Anal. Biochem. 186, 69–73.PubMedCrossRefGoogle Scholar
  2. 2.
    Root, D. D. and Wang, K. (1993) Kinetic silverstaining and calibration of proteins adsorbed to microtiter plates. Anal. Biochem. 209, 354–359.PubMedCrossRefGoogle Scholar
  3. 3.
    Gottlieb, M. and Chavko, M. (1987) Silver staining of native and denatured eucaryotic DNA in agarose gels. Anal. Biochem. 165, 33–37.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 1996

Authors and Affiliations

  • Douglas D. Root
    • 1
  • Kuan Wang
    • 1
  1. 1.Department of Chemistry and BiochemistryUniversity of Texas at Austin

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