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Cetyltrimethylammonium Bromide Discontinuous Gel Electrophoresis of Proteins

Mr-Based Separation of Proteins with Retained Native Activity
  • Robert E. Akins
  • Rocky S. Tuan

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.

Keywords

Sodium Dodecyl Sulfate Native Activity Ammonium Persulfate Coomassie Brilliant Blue Electrophoresis Apparatus 
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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Copyright information

© Humana Press Inc., Totowa, NJ 2002

Authors and Affiliations

  • Robert E. Akins
    • 1
  • Rocky S. Tuan
    • 1
  1. 1.Nemours Biomedical Research Program, A.I. duPont Hospital for ChildrenWilmington

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