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 language | English (US) |
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Pages (from-to) | 691-698 |
Number of pages | 8 |
Journal | Proteomics |
Volume | 1 |
Issue number | 5 |
State | Published - May 2001 |
Externally published | Yes |
Keywords
- Mass spectrometry
- Picomolar protein identification
- Vertical electroelution
ASJC Scopus subject areas
- Genetics
- Molecular Biology