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Free mobility determination by electrophoresis in polyacrylamide containing agarose at a nonrestrictive concentration
Author(s) -
Pospichal Jan,
Vicchio Domenick,
Chrambach Andreas
Publication year - 1991
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.1150120404
Subject(s) - polyacrylamide , agarose , polyacrylamide gel electrophoresis , chromatography , gel electrophoresis of nucleic acids , electrophoresis , chemistry , gel electrophoresis , polymer , agarose gel electrophoresis , analytical chemistry (journal) , dna , biochemistry , polymer chemistry , organic chemistry , enzyme
In the determination of the free mobility, related to the surface net charge, by quantitative gel electrophoresis, the previous arbitrary extrapolation of Ferguson plots from the lowest gel concentrations that give a mechanically stable gel to 0 %T has recently been replaced by measurement of mobilities across that concentration range, using the addition of 0.5 % agarose to polyacrylamide at the various low concentrations in application to a DNA fragment 155 bp in size (Orbán, L. et al ., in preparation). The present study applies that approach to several proteins and DNA fragments smaller than 1300 bp, using 0.4 % agarose in polyacrylamide gels of varying concentration. The intercepts of the plots with the mobility axis provide experimental data by which the free mobility in polyacrylamide gel electrophoresis can be estimated for molecules not significantly retarded in their migration at the agarose concentration admixed to polyacrylamide. Across the gel concentration range below 3 %T, in the presence of agarose, the Ferguson plots of proteins and DNA fragments are convex. It was shown by mass spectrometry that this convex curvature of the plots in the mixed polymer is not significantly due to low polymerization efficiency in the concentration range of liquid polyacrylamide (below 3 %T).