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Electrophoresis. 1997 Mar-Apr;18(3-4):307-16.

Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients.

Author information

1
CEA-Laboratoire de Bioénergétique Cellulaire et Pathologique, UA 2019 DBMS/BECP, Grenoble, France. Thierry@sanrafael.ceng.cea.fr

Abstract

Membrane and nuclear proteins of poor solubility have been separated by high resolution two-dimensional (2-D) gel electrophoresis. Isoelectric focusing with immobilized pH gradients leads to severe quantitative losses of proteins in the resulting 2-D map, although the resolution is usually high. Protein solubility could be improved by using denaturing solutions containing various detergents and chaotropes. Best results were obtained with a denaturing solution containing urea, thiourea, and detergents (both nonionic and zwitterionic). The usefulness of thiourea-containing denaturing mixtures is shown for microsomal and nuclear proteins as well as for tubulin, a protein highly prone to aggregation.

PMID:
9150907
PMCID:
PMC2777268
DOI:
10.1002/elps.1150180303
[Indexed for MEDLINE]
Free PMC Article

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