An improved sodium dodecyl sulfate-polyacrylamide gel electrophoresis system for the analysis of membrane protein complexes.
Membrane protein complexes such as the reaction center complexes of oxygenic photosynthesis or the complex I of mitochondira are composed of many subunit polypeptides. To analyze their polypeptide compositions by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), a wide range of molecular sizes has to be resolved, especially in the low molecular mass range. We have improved the traditional Tris/HCI buffer systems adopting a Tris/2-(N-morpholino)ethanesulfonic acid (MES) buffer system containing 6 M urea. This gel system was used with an 18-24% acrylamide gradient for the separation of polypeptides with molecular masses from below 5 kDa to over 100 kDa. This buffer system can also be applied to the usual uniform concentration of acrylamide gel and also to minislab gels.
Kashino
Y
Y
Department of Life Science, Faculty of Science, Himeji Institute of Technology, Harima Science Garden City, Hyogo, Japan. kashino@sci.himeji-tech.ac.jp
Koike
H
H
Satoh
K
K
D000180:Acrylic Resins; D001426:Bacterial Proteins / Q000032:analysis; D002021:Buffers; D004591:Electrophoresis, Polyacrylamide Gel / Q000379:methods*; D005998:Glycine / Q000031:analogs & derivatives*; D008565:Membrane Proteins / Q000032:analysis; D010455:Peptides / Q000032:analysis; D012967:Sodium Dodecyl Sulfate
D000180:Acrylic Resins; D001426:Bacterial Proteins; D002021:Buffers; D008565:Membrane Proteins; D010455:Peptides; C016680:polyacrylamide gels; D012967:Sodium Dodecyl Sulfate; D005998:Glycine; C100184:tricine