Two-dimensional electrophoresis for the isolation of integral membrane proteins and mass spectrometric identification.
Acrylamide concentration, urea content, and the trailing ion used for sodium dodecyl sulfate (SDS)-gels modify electrophoretic protein mobilities in a protein-dependent way. Varying these parameters we coupled two SDS-gels to a two-dimensional (2-D) electrophoresis system. Protein spots in 2-D gels are dispersed around a diagonal. Hydrophobic proteins are well separated from water-soluble proteins which is the essential advantage of the novel technique. Mass spectrometric identification of previously unaccessible hydrophobic proteins is now possible.
Rais
Isam
I
Institut für Pharmazeutische Chemie, Biozentrum, Universität Frankfurt, Frankfurt am Main, Germany.
Karas
Michael
M
Schägger
Hermann
H
D000818:Animals; D002417:Cattle; D019069:Cell Respiration; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods*; D013058:Mass Spectrometry / Q000379:methods*; D008565:Membrane Proteins / Q000032:analysis* / Q000302:isolation & purification*; D024101:Mitochondrial Proteins / Q000032:analysis* / Q000302:isolation & purification*
D008565:Membrane Proteins; D024101:Mitochondrial Proteins