Direct vertical electroelution of protein from a phastsystem band for mass spectrometric identification at the level of a few picomoles

Zsuzsanna Buzás, Huan Tsung Chang, Nancy E. Vieira, Alfred L. Yergey, Miroslava Stastna, Andreas Chrambach

Research output: Contribution to journalArticlepeer-review

Abstract

An electroelution apparatus prototype of a new design was constructed. In that design, the electric field passes vertically through the protein band located on a horizontal (PhastSystem) minigel polymerized on a net of Gel-Fix (Serva). A simple, home-made apparatus allows for electroelution of protein bands at the level of a few picomoles and their identification, after concentration, by matrix-assisted laser desorption/ionization-time of flight mass spectrometry. The technique is applicable to one-dimensional (1-D) or two-dimensional (2-D) gels of any size, but has been exemplified only by application to 1-D minigels to demonstrate the lower limits of protein load of the method. When in the course of further development of the prototype it will be combined with a modification to two dimensions of the electroelution mechanism under computer control of the high-performance gel electrophoresis apparatus **** (formerly of LabIntelligence), the new design appears uniquely qualified for an automated spot elution from 2-D gels under avoidance of gel sectioning.

Original languageEnglish (US)
Pages (from-to)691-698
Number of pages8
JournalProteomics
Volume1
Issue number5
StatePublished - May 2001
Externally publishedYes

Keywords

  • Mass spectrometry
  • Picomolar protein identification
  • Vertical electroelution

ASJC Scopus subject areas

  • Genetics
  • Molecular Biology

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