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Rheophoretic evaluation of stokes radii in gel electrophoresis

1972; Elsevier BV; Volume: 261; Issue: 1 Linguagem: Inglês

10.1016/0304-4165(72)90324-8

1872-8006

H. WALDMANN-MEYER,

Microfluidic and Bio-sensing Technologies

The effective hydrodynamic radii of native proteins are determined by means of a new analytical method (Rheophoresis), which in one experiment combines the advantages of electrophoretic characterization due to charge differences with information so far only obtainable from chromatography or from a series of electrophoretic experiments in different gel concentrations. The method makes use of the rheophoretic flow, which replaces the water evaporating from biphasic gels (e.g. Sephadex) during quantitative thin-layer electrophoresis. This flow transports all molecules with velocities independent of the net charge but decreasing with retention. Retention is readily evaluated by calculating the rheophoretic flow contribution to the observed migration. The retention coefficients are identical with column chromatographic Vo/Ve ratios and were found to correlate linearly with the Stokes radii of the proteins investigated. The upper separation limit of the gel is directly obtained from this correlation. Rheophoresis stands upon a thoroughly tested theoretical basis. The method is expected to become useful in the quantitative analysis of polymerization and binding equilibria, since interactions between macromolecules or with small ions can be assessed by the simultaneous observation of changes in radius and net charge.