Pulsed field electrophoresis for the separation of protein-sodium dodecyl sulfate-complexes in polyacrylamide gels.
Polyacrylamide gel electrophoresis of proteins was studied using a pulsed-current mode. A new "local field" distribution was used to correct the gel patterns and optimize migration. A corrective field was applied at fixed 2 s intervals to a constant field, inducing a complex relaxation mechanism. Calculated variations in the local field directions decreased the electric strain on the gel during the run, with resultant optimum gel structure. The relaxation mechanism was found to enhance the absolute mobility of proteins with shorter running times compared to constant field gel electrophoresis (CFGE) and other pulsed field techniques. The enhancement of molecular mobility was explored by transverse pore gradient gel electrophoresis. Ferguson curves which exhibited a convex shape in CFGE were linearized by the new pulsed-field method named pulsed oscillatory high-performance electrophoresis (POPE).
Laboratoire de Recherches, Neurosensorielles ORL/Génie Biologique et Médical, Nice, France.
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Houri
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Starita-Geribaldi
D000818:Animals; D016521:Electrophoresis, Gel, Pulsed-Field / Q000295:instrumentation / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel / Q000295:instrumentation / Q000379:methods*; D007202:Indicators and Reagents; D007668:Kidney; D008565:Membrane Proteins / Q000302:isolation & purification; D008871:Microvilli; D008970:Molecular Weight; D011485:Protein Binding; D011506:Proteins / Q000737:chemistry / Q000302:isolation & purification*; D012259:Ribonuclease, Pancreatic / Q000302:isolation & purification; D012709:Serum Albumin / Q000302:isolation & purification; D012967:Sodium Dodecyl Sulfate
D007202:Indicators and Reagents; D008565:Membrane Proteins; D011506:Proteins; D012709:Serum Albumin; D012967:Sodium Dodecyl Sulfate; D012259:Ribonuclease, Pancreatic