Separation of membrane proteins by two-dimensional electrophoresis using cationic rehydrated strips.
Due to their poor solubility during IEF membrane proteins cannot be separated and analyzed satisfactorily with classical 2-DE. A more efficient method for such hydrophobic proteins is the benzyldimethyl-n-hexadecylammonium chloride (16-BAC)/SDS-PAGE, but the corresponding protocol is intricate and time-consuming. We now developed an easy-to-handle electrophoresis method in connection with a novel device which enables reproducible separation of ionic solubilized membrane proteins using individually rehydrated plastic sheet gel strips. These strips are suitable for the first dimension in a 2-D 16-BAC/SDS system and can be handled easily; this is demonstrated by the separation of membrane proteins of human embryonic kidney (HEK293) cells.
Wenge
Birger
B
Institute of Pharmacology and Toxicology, University of Bonn, Bonn, Germany.
Bönisch
Heinz
H
Grabitzki
Julia
J
Lochnit
Günter
G
Schmitz
Brigitte
B
Ahrend
Michael H J
MH
D002460:Cell Line; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D008565:Membrane Proteins / Q000302:isolation & purification*; D015203:Reproducibility of Results; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization
D008565:Membrane Proteins