Ultrathin-layer sodium dodecyl sulfate disc electrophoresis of proteins in the range from 10 to 220 kDa in homogeneous, low-concentrated polyacrylamide gels.
This study describes an ultrathin-layer sodium dodecyl sulfate (SDS) disc electrophoresis in polyacrylamide gels of a thickness of only 150 microm. By use of 2-amino-2-methyl-1,3-propanediol/glycine instead of traditional Tris/HCl buffer in the resolving phase of the gel, proteins with a wide range of molecular sizes (10 kDa to over 220 kDa) are separated in unusually low-concentrated gels (4%T, 3.3%C). 2-Amino-2-methyl-1,3-propanediol in the resolving part of the gel contributes to stabilization of the pH value at 8.8, while glycine improves destacking as well as separation of small proteins from the bulk of stacked SDS. This method combines both the advantages of conventional slab-gel electrophoresis and capillary gel electrophoresis. It is easy to apply and well suited for all further miniaturization attempts.
Maly
I Piotr
IP
Institute of Anatomy, University of Basel, Basel, Switzerland. i-p.maly@unibas.ch
Crotet
Valérie
V
Toranelli
Mireille
M
D000818:Animals; D002021:Buffers; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods* / Q000592:standards; D005782:Gels; D008832:Microchemistry / Q000295:instrumentation / Q000379:methods; D008970:Molecular Weight; D011506:Proteins / Q000302:isolation & purification*; D051381:Rats
D002021:Buffers; D005782:Gels; D011506:Proteins