The sieving of spheres during agarose gel electrophoresis: Quantitation and modeling

Gary A. Griess, Elena T. Moreno, Richard A. Easom, Philip Serwer

Research output: Contribution to journalArticle

75 Scopus citations

Abstract

By use of agarose gel electrophoresis, the sieving of spherical particles in agarose gels has been quantitated and modeled for spheres with a radius (R) between 13.3 and 149 mm. For quantitation, the electrophoretic mobility has been determined as a function of agarose percentage (A). Because a previously used model of sieving [D. Rodbard and A. Chrambach (1970) Proc. Natl. Acad. Sci. USA 65, 970–977] was found incompatible with some of these data, alternative models have been tested. By use of an underivatized agarose, two models, both based on the assumption of a single effective pore radius (PE) for each A, were found to yield PE values that were independent of R and that were in agreement with values of PE obtained independently (PE = 118 nm × A−0.74): sieving by altered hydrodynamics in a cylindrical tube of radius, PE, and sieving by steric exclusion from a circular hole of radius, PE. The same analysis applied to a 6.5% hydroxyethylated commercial agarose yileded a steeper PE vs A plot and also agreement of the above tow models with the data. The PE vs A plot was significantly altered by both further hydroxyethylation and factors that cause variation in the electro‐osmosis found in commercial agarose.

Original languageEnglish (US)
Pages (from-to)1475-1484
Number of pages10
JournalBiopolymers
Volume28
Issue number8
DOIs
StatePublished - Aug 1989

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Biomaterials
  • Organic Chemistry

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