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

Thierry Rabilloud (1) , C. Adessi (2) , A. Giraudel (3) , J. Lunardi (1)

1 BECP - Bioénergétique Cellulaire et Pathologique
2 Développement de la protéomique comme outil d'investigation fonctionelle et d'annotation des génomes
3 Organisation Fonctionnelle du Cytosquelette

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.

Mots clés: two-dimensional electrophoresis; protein solubilization; membrane proteins; nuclear proteins

Domaines: Génomique, Transcriptomique et Protéomique [q-bio.GN]

Thierry Rabilloud, C. Adessi, A. Giraudel, J. Lunardi. Improvement of the solubilization of proteins in two-dimensional electrophoresis with immobilized pH gradients.. Electrophoresis, 1997, 18 (3-4), pp.307-16. ⟨10.1002/elps.1150180303⟩. ⟨hal-00118181⟩