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Protein Detection in Polyacrylamide Gel Electrophoresis

  • Bonnie S. Dunbar

Abstract

An investigator must take several factors into account for the process of staining and recording the location of bands after electrophoresis (Wilson, 1979, 1983). These should include (1) the selection and availability of an appropriate dye of satisfactory quality that will give adequate sensitivity; (2) the selection of a solvent that will provide a suitable dye concentration, solubility, and stability; (3) the formation of a stable dye—protein complex; (4) the easy removal of excess free dye for easy detection of stained proteins; and (5) storage of gels for further reference or for permanent recording by photography or analysis by computer, densitometry, or spectrophotometry.

Keywords

Sodium Dodecyl Sulfate Protein Detection Silver Stain Acid Blue Silver Staining Method 
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.

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References

  1. Carson, S. D., 1977, Hydrazinocridine staining of proteins and glycoproteins in polyacrylamide gels, Anal Biochem. 78:428–435.PubMedCrossRefGoogle Scholar
  2. Dunbar, B. S., Dudkiewicz, A. B., and Bundman, D. S., 1985, Proteolysisofspecific porcine zona pellucida glycoproteins by boar acrosin, Biol. Reprod. 32:619–630.PubMedCrossRefGoogle Scholar
  3. Hames, B. D., 1981, An introduction to polyacrylamide gel electrophoresis, in: Gel Electrophoresis of Proteins: A Practical Approach (B. D. Hames and D. Rickwood, eds.), pp. 1–91, IRL Press, Oxford, England.Google Scholar
  4. Hartman, B. K., and Udenfriend, S., 1969, A method for immediate visualization of proteins in acrylamide gels and its use for preparation of antibodies to enzymes, Anal. Biochem. 30:391–394.PubMedCrossRefGoogle Scholar
  5. Higgins, R. C., and Dahmus, M. E., 1979, Rapid visualization of protein bands in preparative SDS-polyacrylamide gels, Anal. Biochem. 93:257–260.PubMedCrossRefGoogle Scholar
  6. Hitchcock, P. J., and Brown, T. M., 1983, Morphological heterogeneity among Salmonella lipopolysaccharide chemotypes in silver stained polyacrylamide gels, J. Bacteriol. 154:269–277.PubMedGoogle Scholar
  7. Jackowski, G., and Liew, C. C., 1980, Fluorescamine staining of non-histone chromatin proteins as revealed by two-dimensional polyacrylamide gel electrophoresis, Anal. Biochem. 102:321–325.PubMedCrossRefGoogle Scholar
  8. Marshall, T., and Williams, K. M., 1984, Artifacts associated with 2-mercaptoethanol upon high-resolution two-dimensional electrophoresis, Anal. Biochem. 139:502–505.PubMedCrossRefGoogle Scholar
  9. Matheka, H. D., Enzmann, P. J., Bachrach, H. L., and Migl, B., 1977, Anal. Biochem. 81:9–000.PubMedCrossRefGoogle Scholar
  10. Merril, C. R., and Goldman, D., 1984, Detection of polypeptides in two-dimensional gels using silver staining, in: Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications (J. E. Celis and R. Bravo, eds.), pp. 111–126, Academic Press, New York.Google Scholar
  11. Merril, C. R., Goldman, D., and Van Keuren, M. L., 1982, Simplified silverproteindetection and image enhancement methods in polyacrylamide gels, Electrophoresis 3:17–23.CrossRefGoogle Scholar
  12. Merril, C. R., Switzer, R. C., and Van Keuren, M. L., 1979, Trace polypeptides in cellular extracts and human body fluids detected by two-dimensional electrophoresis and highly sensitive silver stain, Proc. Natl. Acad. Sci. USA 76:4335–4339.PubMedCrossRefGoogle Scholar
  13. Nelles, L. P., and Bamburg, J. R., 1976, Rapid visualization of protein-dodecyl sulfate complexes in polyacrylamide gels, Anal. Biochem. 73:522–531.PubMedCrossRefGoogle Scholar
  14. Ochs, D., 1983, Protein contaminations of sodium dodecyl sulfate polyacrylamide gels, Anal. Biochem. 135:470–474.PubMedCrossRefGoogle Scholar
  15. Ochs, D. C., McConkey, E. H., and Sammons, D. W., 1981, Silver stains for protein in polyacrylamide gels: A comparison of six methods, Electrophoresis 2:304–307.CrossRefGoogle Scholar
  16. Ragland, W. L., Pace, J. L., and Kemper, D. L., 1974, Fluorometric scanning of fluores- camine-labeled proteins in polyacrylamide gels, Anal. Biochem. 59:24–33.PubMedCrossRefGoogle Scholar
  17. Sammons, D. W., Adams, L. D., and Nishizawa, E. E., 1981, Ultrasensitive silver-based color staining of polypeptides in polyacrylamide gels, Electrophoresis 3:135–141.CrossRefGoogle Scholar
  18. Sammons, D. W., Adams, L. D., Vidmar, T. J., Hatfield, C. A., Jones, D. H., Chuba, P. J., and Crooks, S. W., 1984, Applicability of color silver stain (Gelcode(r) System) to protein mapping with two-dimensional gel electrophoresis, in: Two-Dimensional Gel Electrophoresis of Proteins: Methods and Applications (J. E. Celis and R. Bravo, eds.), pp. 111–126, Academic Press, New York.Google Scholar
  19. Schetters, H., and McLeod, B., 1979, Simultaneous isolation of major viral proteins in one step, Anal. Biochem. 98:329–334.PubMedCrossRefGoogle Scholar
  20. 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.PubMedCrossRefGoogle Scholar
  21. Takagi, T., Kubo, K., and Isemura, T., 1977, Simple visualization of protein bands in SDS-polyacrylamide gel electrophoresis by the insoluble complex formation between SDS and a cationic surfactant, Anal. Biochem. 79:104–109.PubMedCrossRefGoogle Scholar
  22. Tjissen, P., and Kurstak, E., 1979, A simple and sensitive method for the purification and peptide mapping of proteins solubilized from densonucleosis virus with sodium dodecyl sulfate, Anal. Biochem. 99:97–104.CrossRefGoogle Scholar
  23. Wilson, C. M., 1979, Studies and critique of amido Black 10B, Coomassie Blue R, and Fast Green FCF as stains for proteins after polyacrylamide gel electrophoresis, Anal. Biochem. 9:263–278.CrossRefGoogle Scholar
  24. Wilson, C. M., 1983, Staining of proteins on gels: Comparisons of dyes and procedures, Methods Enzymol. 91:236–247.PubMedCrossRefGoogle Scholar
  25. Wray, W., Boulikas, T., Wray, V. P., and Hancock, R., 1981, Silver staining of proteins in polyacrylamide gels, Anal. Biochem. 118:197–203.PubMedCrossRefGoogle Scholar

Copyright information

© Plenum Press, New York 1987

Authors and Affiliations

  • Bonnie S. Dunbar
    • 1
  1. 1.Baylor College of MedicineHoustonUSA

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