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

Proteomics. 2001 May;1(5):691-8. doi: 10.1002/1615-9861(200104)1:5<691::AID-PROT691>3.0.CO;2-7.

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.

MeSH terms

  • Electrophoresis, Polyacrylamide Gel / methods*
  • Microchemistry / methods*
  • Serum Albumin, Bovine / chemistry*
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization / methods*

Substances

  • Serum Albumin, Bovine