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Issue 12, 2014
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Protein/peptide purification by three-well OFFGEL electrophoresis with immobilized ultra narrow pH gradient gels

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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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Supplementary files

Article information


Submitted
09 Jan 2014
Accepted
07 Apr 2014
First published
07 Apr 2014

Anal. Methods, 2014,6, 3995-4002
Article type
Paper
Author version available

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

E. Tobolkina, F. Cortés-Salazar, L. Qiao and H. H. Girault, Anal. Methods, 2014, 6, 3995
DOI: 10.1039/C4AY00088A

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