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Cetyltrimethylammonium Bromide Discontinuous Gel Electrophoresis of Proteins

M r -Based Separation of Proteins with Retained Native Activity

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Book cover The Protein Protocols Handbook

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Abstract

This chapter describes a novel method of electrophoresis that allows the fine separation of proteins to be carried out with the retention of native activity. The system combines discontinuous gel electrophoresis in an arginine/N-Tris (hydroxymethyl) methylglycine) (Tricine) buffer with sample solubilization in cetyltrimethylammonium bromide (CTAB). Because the components that distinguish this system are CTAB, arginine, and Tricine and because CTAB is a cationic detergent, we refer to this method as CAT gel electrophoresis (1,2). Proteins separated on CAT gels appear as discrete bands, and their mobility is a logarithmic function of M r across a broad range of molecular weights. After CAT electrophoresis, many proteins retain high enough levels of native activity to be detected, and gel bands may be detected by both M r and protein-specific activities. In this chapter, we provide a description of the procedures for preparing and running CAT gels. We also provide some technical background information on the basic principles of CAT gel operation and some points to keep in mind when considering the CAT system.

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Author information

Authors and Affiliations

  1. Department of Medical Cell Biology, Nemours Research Program, AI DuPont Institute, Wilmington, DE

    Robert E. Akins

  2. Department of Orthopedic Surgery, Thomas Jefferson University, Philadelphia, PA

    Rocky S. Tuan

Authors
  1. Robert E. Akins
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  2. Rocky S. Tuan
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Editor information

Editors and Affiliations

  1. University of Hertfordshire, Hatfield, UK

    John M. Walker

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© 1996 Humana Press Inc., Totowa, NJ

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Akins, R.E., Tuan, R.S. (1996). Cetyltrimethylammonium Bromide Discontinuous Gel Electrophoresis of Proteins. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1007/978-1-60327-259-9_13

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  • DOI: https://doi.org/10.1007/978-1-60327-259-9_13

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-338-2

  • Online ISBN: 978-1-60327-259-9

  • eBook Packages: Springer Book Archive

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