TY - JOUR
T1 - The sieving of spheres during agarose gel electrophoresis
T2 - Quantitation and modeling
AU - Griess, Gary A.
AU - Moreno, Elena T.
AU - Easom, Richard A.
AU - Serwer, Philip
PY - 1989/8
Y1 - 1989/8
N2 - 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.
AB - 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.
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U2 - 10.1002/bip.360280811
DO - 10.1002/bip.360280811
M3 - Article
C2 - 2752101
AN - SCOPUS:0024720506
SN - 0006-3525
VL - 28
SP - 1475
EP - 1484
JO - Biopolymers
JF - Biopolymers
IS - 8
ER -