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Analysis of convex Ferguson plots in agarose gel electrophoresis by empirical computer modeling
Author(s) -
Tietz Dietmar,
Chrambach Andreas
Publication year - 1986
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.1150070602
Subject(s) - agarose , radius , agarose gel electrophoresis , fiber , chemistry , electrophoresis , chromatography , hydrodynamic radius , analytical chemistry (journal) , materials science , polymer , biochemistry , organic chemistry , computer security , computer science , copolymer , gene
Abstract Agarose gel electrophoresis has been shown to give rise to non‐linear plots of log (mobility) vs. gel concentration of spherical viruses (Serwer,[1]) and cellular vesicles (Gottlieb et al. [2]). This finding also applies to proteins as shown in this study. Considering that in the non‐linear plot, the slope becomes a function of gel concentration, it is possible to determine particle properties and gel parameters by a modification of the conventional method derived from the Ogston theory for long‐fiber gels. This treatment shows: (a) In application to data obtained from gel electrophoresis (0.4 to 1.6 % agarose) of viruses (13 to 42 nm radius) and with increasing gel concentration: (i) an increase of apparent total fiber length per g agarose matrix, (ii) a reduction of apparent fiber radius and (iii) a constant fiber volume (per g matrix material) of the agarose fiber. The values of the fiber radii identify the fibers as agarose supercoils (or aggregates of it) with a radius of 20–55 nm. (b) In application to data obtained from gel electrophoresis (1.2 to 8 % agarose) of proteins (1.7 to 5.8 nm radius): a fiber volume indicative of additional sieving by the agarose double helix (of known radius of 0.5–2 nm). This is in agreement with a previous suggestion by Serwer [1] that proteins are able to penetrate into the double‐helical network of which the agarose supercoil consists. (c) The likelihood of a continuous transition from case (a) to case (b).