Advertisement

Electroelution of proteins from bands in gel electrophoresis without gel sectioning for the purpose of protein transfer into mass spectrometry: Elements of a new procedure

Authors

  • Huan-Tsung Chang,

    1. Section on Macromolecular Analysis, Laboratory of Cellular and Molecular Biophysics, NICHD, NIH, Bethesda, MD, USA
    2. Department of Chemistry, National Taiwan University, Taipei, Taiwan
    Search for more papers by this author
  • Alfred L. Yergey,

    1. Section on Mass Spectrometry and Metabolism, Laboratory of Cellular and Molecular Biophysics, NICHD, NIH, Bethesda, MD, USA
    Search for more papers by this author
  • Andreas Chrambach

    Corresponding author
    1. Section on Macromolecular Analysis, Laboratory of Cellular and Molecular Biophysics, NICHD, NIH, Bethesda, MD, USA
    • Section on Macromolecular Analysis, Laboratory of Cellular and Molecular Biophysics, NICHD, Bldg.10, Rm. 9D50, NIH, Bethesda MD 20892-1580, USA, Fax: +301-402-0263
    Search for more papers by this author

Abstract

Electroelution of protein bands from a gel has advantages over the competitive common technique requiring gel sectioning with respect to yield, speed and the potential for computer-controlled application to multicomponent two-dimensional (2-D) gels. The electroelution design for the commercial high-performance gel electrophoresis (HPGE) apparatus represented the most advanced technique to date until the recent discontinuation of its production. The present report serves to summarize the necessary design elements for the purpose of renewing and further developing the electroelution technique. A rudimentary technique is presented by which the electroeluate is collected in a glass tube superimposed on a reversibly stained gel band and connected to an anolyte reservoir. Although the stain used is insufficiently sensitive, the technique allowed for the qualitative verification of its usefulness in the transfer of the electroeluate into mass spectrometry.

Ancillary