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Protein/peptide purification by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels†
Elena Tobolkina,Fernando Cortés-Salazar,Liang Qiao,Hubert H. Girault
Analytical Methods Pub Date : 04/07/2014 00:00:00 , DOI:10.1039/C4AY00088A
Abstract

Purification and desalting of protein and peptide samples by 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 proteins and peptides according to their isoelectric points 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.

Graphical abstract: Protein/peptide purification by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels
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