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

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.

Cite this paper

@article{Buzs2001DirectVE, title={Direct vertical electroelution of protein from a PhastSystem band for mass spectrometric identification at the level of a few picomoles.}, author={Zsuzsanna Buz{\'a}s and H. T. Chang and Nancy E. Vieira and Alfred L Yergey and Miroslava Stastn{\'a} and Andreas Chrambach}, journal={Proteomics}, year={2001}, volume={1 5}, pages={691-8} }