Gel electrophoresis in the presence of soluble, aqueous polymers: horizontal sodium dodecyl sulfate-polyacrylamide gels.
Because little is known about the use of aqueous polymers in polyacrylamide gel electrophoresis, we undertook a feasibility study that enables the discontinuous Laemmli-formulated system of sodium dodecyl sulfate (SDS)-polyacrylamide gel electrophoresis to be performed in a horizontal format by the addition of large-sized aqueous polymers (i.e., dextrans and methylcelluloses). We studied four parameters: the cross-linking agent (bisacrylamide vs AcrylAide) and the polymer concentration, nature, and size. Three concentrations of each polymer were used. The best differentiation between the standard markers and the sharpest bands were obtained using concentrations of 2.5 and 0.06% (w/v) for Dextran T-500 and methylcellulose 4000, respectively. There was no predictable pattern to the variation in the plots of log Mr vs Rf caused by varying the concentration and length of the dextrans; however, the methylcellulose patterns suggest that gel viscosity is important. The results suggest that the combination of 0.06% methylcellulose 4000 polymers with bisacrylamide is a convenient and inexpensive means of performing flatbed Laemmli SDS-polyacrylamide gel electrophoresis.
Gersten
D M
DM
Department of Pathology, Georgetown University Medical Center, Washington, D.C. 20007.
Kimball
H
H
Bijwaard
K E
KE
D003911:Dextrans; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D008747:Methylcellulose; D011108:Polymers; D011506:Proteins / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate; D012995:Solubility; D014867:Water
D003911:Dextrans; D011108:Polymers; D011506:Proteins; D014867:Water; D012967:Sodium Dodecyl Sulfate; D008747:Methylcellulose