SDS disc electrophoresis of proteins in homogeneous, low-concentrated polyacrylamide gels.
In the attempt to separate in a single gel run low- and high-molecular-weight proteins, we present here a multiphasic buffer system designed for this purpose. It avoids the continuous stacking of SDS as it occurs in the 'classical' SDS-PAGE. The system allows complete stacking and destacking of proteins in the 3.5-250 kDa range at acrylamide concentrations as low as 4.5% T (total acrylamide concentration in %) and 2.6% C (degree of cross-linking in %). Taurine is used as the trailing ion in the cathode buffer and in the resolving zone of the gel, and two different counterions (Tris and imidazole) in the stacking zone. The gel system is easy to prepare and, due to the very low acrylamide concentrations, it is ideal for analytical as well as for preparative tasks.
Maly
I Piotr
IP
Section of Functional Neuroanatomy, Institute of Anatomy, DKBW, University of Basel, Basel, Switzerland. i-p.maly@unibas.ch
Nitsch
Cordula
C
D000180:Acrylic Resins / Q000737:chemistry*; D000818:Animals; D001923:Brain Chemistry; D002021:Buffers; D004589:Electrophoresis, Disc / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D006863:Hydrogen-Ion Concentration; D007202:Indicators and Reagents / Q000592:standards; D008099:Liver / Q000737:chemistry; D051379:Mice; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D012015:Reference Standards; D013654:Taurine; D014325:Tromethamine
D000180:Acrylic Resins; D002021:Buffers; D007202:Indicators and Reagents; D011506:Proteins; C016680:polyacrylamide gels; D014325:Tromethamine; D013654:Taurine