Premium
Sieving of ionic constituents across moving boundaries in gel electrophoresis
Author(s) -
Orbán László,
Fawcett John S.,
Tietz Dietmar,
Chrambach Andreas
Publication year - 1989
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.1150100407
Subject(s) - agarose , polyacrylamide , chemistry , chloride , chromatography , ionic strength , analytical chemistry (journal) , electrophoresis , polyacrylamide gel electrophoresis , aqueous solution , biochemistry , polymer chemistry , organic chemistry , enzyme
The representative β‐hydroxyethylmorpholinium‐chloride‐bicinate moving boundary with a trailing ion net mobility relative to Na + of 0.41, detected by precipitation of chloride with silver nitrate, exhibits a decreasing chloride mobility at increasing polyacrylamide gel concentrations from 3.5 to 45 %T, 5 %C Bis . This decrease, largely due to an increase of field strength at constant current, is described by a convex plot of log (mobility) vs. %T (Ferguson plot) and signifies that chloride/bicinate are sieved by the gel. In agarose gels, the same plot of mobility vs. gel concentration is constant below 7 % gel concentration, since in those gels field strength and migration rate remain the same within that gel concentration range. Both in polyacrylamide and in agarose gels the displacement rate of the chloride‐bicinate boundary as a function of the time of electrophoresis or distance migrated remains invariant within 15 %. The plot of log(mobility) vs. gel concentration extrapolated to 0 %T is 5.85 and 5.41 (10 −5 cm 2 s −1 V −1 ) for polyacrylamide and for agarose (SeaKem HGT‐P, FMC) gels, respectively. The slightly decreased mobility intercept at 0 %T for agarose is presumably due either to the electroendosmotic properties of agarose HGT‐P and/or failure to Sufficiently take into account the flattening of the Ferguson plot in the polyacrylamide concentration range below 3 % in which a transition from a gel to a fluid (sol) medium takes place.