Proteins/peptides purification by a three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels

Purification and desalting of protein and peptide samples by a three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels is proposed as a fast preparative strategy for proteomics. The gist of this strategy is to separate the proteins and peptides according to their isoelectric point and to isolate those of a given pI value equal to the mean pH value of the gel. The present approach has been demonstrated both on protein mixtures and a digested Escherichia coli protein extract. UV-Vis spectroscopy, MALDI-MS, SDS-PAGE and LC-MS/MS were employed for the quantitative and qualitative characterization of the separation results. The electrophoretic methodology has been simulated by finite element methods.


Published in:
Analytical Methods, 6, 3995-4002
Year:
2014
Publisher:
Cambridge, Royal Society of Chemistry
ISSN:
1759-9660
Laboratories:




 Record created 2014-04-07, last modified 2018-03-17

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