COMPARISON OF THE EFFECTIVITY OF 2 DIFFERENT REHYDRATION SOLUTIONS ONTHE SOLUBILIZATION OF PROTEINS SEPARATED BY 2-DIMENSIONAL ELECTROPHORESIS

An immediate advance of two-dimensional 2(DE) gel electrophoresis with immobilized pH gradients is the practical feasibility of setting up v ery narrow pH gradients in the most regions of the pH scale, with resu ltant dramatic improvement in resolution for some applications. Howeve r, when less soluble proteins are analyzed by this technique, quantita tive protein losses are observed. It was shown that protein solubility could be improved by varying the detergents and chaotropes used for r ehydration of the immobiline strips. Our results confirmed the high ef fectiveness of a modified urea/thiourea/sulfobetaines rehydration buff er in the solubilization of proteins occurring, especially, in acidic area of 2-DE gels in which the presence of cytoskeletal proteins can b e anticipated. It was also found that proteins requiring a higher amou nt of chaotropic substances to enter imobiline strips could be detecte d using a standard rehydration solution and they are lost when the ure a concentration is diminished.