Direct vertical electroelution of protein from a PhastSystem band for mass spectrometric identification at the level of a few picomoles
✍ Scribed by Zsuzsanna Buzás; Huan-Tsung Chang; Nancy E. Vieira; Alfred L. Yergey; Miroslava Stastna; Andreas Chrambach
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 112 KB
- Volume
- 1
- Category
- Article
- ISSN
- 1615-9853
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✦ Synopsis
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/ionizationtime 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.