1. CONFECCIÓN DE GELES DE POLIACRILAMIDA EN CONDICIONES DE LABORATORIO PARA FOCALIZACIÓN ISOELECTRICA.
- Author
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Manrique Suárez, Viana, Higginson Clarke, David, and Riverón Rojas, Ana Maria
- Subjects
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PROTEINS , *ISOELECTRIC focusing , *POLYACRYLAMIDE gel electrophoresis , *COLLOIDS , *ACRYLAMIDE , *ION exchange (Chemistry) - Abstract
Bidimensional electroforesis is the method more employed for the analysis of complex mixtures of extracted proteins of cells, tissue or other biological samples. Using this technique the proteins are separated according to two independent properties in two steps: a first dimension where these are fractioned on the base of their isoelectric point through isoelectric focusing and a second dimension in which the previously separate proteins are fractioned based on their dimensions through SDS poliacrylamide gel electrophoresis. Isoelectric focusing is generally carried out using expensive commercial gels (e.g. Immobiline DryStrip gel). A home mademethod was developed to produce gels useful for IEF. 30% T Acrylamide and 8M Urea solutions were deionized using mixed bed ion exchangers. The quality of the deionization processes was evaluated through the variations of solutions´ conductivities of both reagents. Acrylamide deionization was also evaluated through variations in the pH and the Absorbance at 290 nm. The obtained data were processed statistically with STATISTIC 6.0.Two electrophoresis formats, circular gels and horizontal gels were tested. The applied sample was human plasma delipidated with dithiothreitol and Tween-80 for 15 min. There were significant differences in the values of conductivity before and after deionización, (p <0.05), for both solutions. The pattern of bands was obtained satisfactorily for the two systems when deioniozed solutions were used. These results allow to conclude that gels prepared with the developed system substitute satisfactorily the commercial gels for identical purposes. By this way we contribute to decrease the 10/90 gap (only 10% of the world funds for health research is used in the investigation of 90% of the world problems of health), where the reduction of the costs of the analytic methods is among the strategies outlined by the Global Forum for Investigations in Health for teh 2003-2005 period. [ABSTRACT FROM AUTHOR]
- Published
- 2005