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Protein Blotting and Detection pp 115–128Cite as

Cationic electrophoresis and eastern blotting

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 536))

Summary

Denaturing, discontinuous electrophoresis in the presence of SDS has become a standard method for the protein scientist. However, there are situations where this method produces suboptimal results. In these cases, electrophoresis in the presence of positively charged detergents like cetyltrimethylammonium bromide may work considerably better. Methods for electrophoresis, staining, and blotting of such gels are presented.

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References

  1. Laemmli, U. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.

    Article  PubMed  CAS  Google Scholar 

  2. Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretictransfer of proteins from polyacrylamide gels to nitrocellulose sheets: Procedure and some applications. Proc Natl Acad Sci USA 76, 4350–4354.

    Article  PubMed  CAS  Google Scholar 

  3. Dunn, S. (1986) Effects of the modifica­-tion of transfer buffer composition and the renaturation of proteins in gels on the recognition of proteins on western blots by monoclonal antibodies. Anal Biochem 157, 144–153.

    Article  PubMed  CAS  Google Scholar 

  4. Shirahama, K., Tsujii, K., and Takagi, T. (1974) Free boundary electrophoresis of sodium dodecyl sulphate-protein polypeptide complexes with special reference to SDS-polyacrylamide gel electrophoresis. J Biochem 75, 309–319.

    PubMed  CAS  Google Scholar 

  5. Rabilloud, T., Girardot, V., and Lawrence, J.-J. (1996) One- and two dimensional histone separations in acidic gels: Usefulness of methylene blue-driven photopolymerisation. Electrophoresis 17, 67–73.

    Article  PubMed  CAS  Google Scholar 

  6. Panyim, S., Thitipongpanich, R., and Supatimusro, D. (1977) A simplified gel electrophoretic system and its validity for molecular weight determinations of protein–cetyltrimethylammonium complexes. Anal Biochem 81, 320–327.

    Article  PubMed  CAS  Google Scholar 

  7. Eley, M., Burns, P., Kannapell, C., and ­Campbell, P. (1979) Cetyltrimethylammonium bromide polyacrylamide gel electrophoresis: Estimation of protein subunit molecular weights using cationic detergents. Anal Biochem 92, 411–419.

    Article  PubMed  CAS  Google Scholar 

  8. MacFarlane, D. (1983) Use of ­benzyldimethyl-n-hexadecylammonium chloride (“16-BAC”), a cationic detergent, in an acidic polyacrylamide gel electrophoresis system to detect base labile protein methylation in intact cells. Anal Biochem 132, 231–235.

    Article  PubMed  CAS  Google Scholar 

  9. Mócz, G., and Bálint, M. (1984) Use of cat­ionic detergents for polyacrylamide gel electrophoresis in multiphasic buffer systems. Anal Biochem 143, 283–292.

    Article  PubMed  Google Scholar 

  10. Akin, D., Shapira, R., and Kinkade, J., Jr. (1985) The determination of molecular weights of biologically active proteins by cetyl­trimethylammonium bromide-polyacryl­amide gel electrophoresis. Anal Biochem 145, 170–176.

    Article  PubMed  CAS  Google Scholar 

  11. Akins, R., Levin, P., and Tuan, R. (1992) Cetyltrimethylammonium bromide discontinuous electrophoresis: Mr-based separation of proteins with retention of enzymatic activity. Anal Biochem 202, 172–178.

    Article  PubMed  CAS  Google Scholar 

  12. Hartinger, J., Stenius, K., Högemann, D., and Jahn, R. (1996) 16-BAC/SDS-PAGE: A two-dimensional gel electrophoresis system suitable for the separation of integral membrane proteins. Anal Biochem 240, 126–133.

    Article  PubMed  CAS  Google Scholar 

  13. Ornstein, L. (1962) Disk electrophoresis: I. Background and theory. Ann N Y Acad Sci 121, 321–351.

    Article  Google Scholar 

  14. Jovin, T. (1973) Multiphasic zone electrophoresis. I. Steady-state moving-boundary systems formed by different electrolyte combinations. Biochemistry 12, 871–879.

    Article  PubMed  CAS  Google Scholar 

  15. Jovin, T. (1973) Multiphasic zone electro­phoresis. II. Design of integrated dis­continuous buffer systems for analytical and preparative fractionation. Biochemistry 12, 879–890.

    Article  PubMed  CAS  Google Scholar 

  16. Jovin, T. (1973) Multiphasic zone electrophoresis. III. Further analysis and new forms of discontinuous buffer systems. Biochemistry 12, 890– 898.

    Article  PubMed  CAS  Google Scholar 

  17. Jovin, T. (1973) Multiphasic zone electrophoresis. IV Design and analysis of discontinuous buffer systems with a digital computer. Ann N Y Acad Sci 209, 477–496.

    Article  PubMed  CAS  Google Scholar 

  18. Chrambach, A. (1985) The practice of quantitative gel electrophoresis. VCH, Weinheim.

    Google Scholar 

  19. Buxbaum, E. (2003) Cationic electrophoresis and electrotransfer of membrane glycoproteins. Anal Biochem 314, 70–76.

    Article  PubMed  CAS  Google Scholar 

  20. Pritchard, D., Crawford, C., Duce, I., and Behnke, J. (1985) Antigen stripping from the nematode epicuticle using the cationic detergent cetyltrimethylammonium bromide (CTAB). Parasite Immunol 7, 575–585.

    Article  PubMed  CAS  Google Scholar 

  21. Freedman, D., Nutman, T., and Ottesen, E. (1988) Enhanced solubilization of immunoreactive proteins from Brugia malayi adult parasites using cetyltrimethylammonium bromide. Exp Parasitol 65, 244–250.

    Article  PubMed  CAS  Google Scholar 

  22. Maki, K., Sagara, J., and Kawai, A. (1991) A cationic detergent, cetyltrimethylammonium bromide (CTAB), selectively dissociates the intermediate filament of the fibroblast. Biochem Biophys Res Commun 175, 768–774.

    Article  PubMed  CAS  Google Scholar 

  23. Smith, I., Cromie, R., and Stainsby, K. (1988) Seeing gel wells well. Anal Biochem 169, 370–371.

    Article  PubMed  CAS  Google Scholar 

  24. Lazar, J., and Taub, F. (1992) A highly sensitive method for detecting peroxidase in situ hybridization or immunohistochemical assays. In Nonradioactive labeling and detection of biomolecules, Kessler, C., ed., pp. 135–142. Springer, Berlin.

    Google Scholar 

  25. Rodbard, D., and Chrambach, A. (1971) Estimation of molecular radius, free mobility, and valence using polyacrylamide gel electrophoresis. Anal Biochem 40, 95–134.

    Article  PubMed  CAS  Google Scholar 

  26. Stellwagen, N. (1998) Apparent pore size of polyacrylamide gels. Electrophoresis 19, 1542–1547.

    Article  PubMed  CAS  Google Scholar 

  27. Heukeshoven, J., and Dernick, R. (1988) Improved silver staining procedure for fast staining in phastsystem development unit I. Staining of sodium dodecyl sulfate gels. Electrophoresis 9, 28–32.

    Article  PubMed  CAS  Google Scholar 

  28. Merril, C., Goldman, D., Sedman, S., and Ebert, M. (1981) Ultrasensitive stain for proteins in polyacrylamide gels shows regional variations in cerebrospinal fluid proteins. Science 211, 1437–1438.

    Article  PubMed  CAS  Google Scholar 

  29. Segrest, J., and Jackson, R. (1972) Molecular weight determination of glycoproteins by polyacrylamide gel electrophoresis in sodium dodecyl sulfate. Methods Enzymol. 28, 54–63.

    Article  Google Scholar 

  30. Thornton, D., Holmes, D., Sheehan, J., and Carlstedt, I. (1989) Quantitation of mucus glycoproteins blotted onto nitrocellulose membranes. Anal Biochem 182, 160–164.

    Article  PubMed  CAS  Google Scholar 

  31. Hancock, K., and Tsang, V. (1983) India ink staining of proteins on nitrocellulose paper. Anal Biochem 133, 157–162.

    Article  PubMed  CAS  Google Scholar 

  32. Hassan, J., Feighery, C., and Whalan, A. (1987) Staining molecular weight markers on nitrocellulose using Ponceau S. J Clin Lab Immunol 24, 104.

    PubMed  CAS  Google Scholar 

  33. Moore, M., and Viselli, S. (2000) Staining and quantification of proteins transferred to polyvinylidene difluoride membranes. Anal Biochem 279, 241–242.

    Article  PubMed  CAS  Google Scholar 

  34. Leong, M., and Fox, G. (1990) Luminescent detection of immunodot and Western blot. Methods Enzymol 184, 442–451.

    Article  PubMed  CAS  Google Scholar 

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Acknowledgements

This work was supported in part by Kuwait University grant MPB029.

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Authors and Affiliations

  1. Department of Biochemistry, Ross University School of Medicine, Box 266, Portsmouth Campus, Roseau, Commonwealth of Dominica, West Indies

    Engelbert Buxbaum

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  1. Engelbert Buxbaum
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Correspondence to Engelbert Buxbaum .

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Editors and Affiliations

  1. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    Biji T. Kurien

  2. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    R. Hal Scofield

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Buxbaum, E. (2009). Cationic electrophoresis and eastern blotting . In: Kurien, B., Scofield, R. (eds) Protein Blotting and Detection. Methods in Molecular Biology, vol 536. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-542-8_14

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  • DOI: https://doi.org/10.1007/978-1-59745-542-8_14

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-73-2

  • Online ISBN: 978-1-59745-542-8

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