How to bring the "unseen" proteome to the limelight via electrophoretic pre-fractionation techniques.
The present review reports a panoply of electrophoretic methods as pre-fractionation tools in proteomic investigations in preparation for mass spectrometry or two-dimensional electrophoresis map analysis. Such electrophoretic pre-fractionation protocols include all those electrokinetic methodologies which are performed in free solution, most of them relying on isoelectric focusing steps (although some approaches based on gels and granulated media are also discussed). Devices associated with electrophoretic separations are multi-chamber apparatuses, such as the multi-compartment electrolyzers equipped with either isoelectric membranes or with isoelectric beads, Off-Gel electrophoresis in a multi-cup device and the Rotofor, an instrument also based on a multi-chamber system but exploiting the conventional technique of carrier-ampholyte-focusing. Other free-flow systems, as well as miniaturized chambers, are also described.
Righetti
Pier Giorgio
PG
Department of Industrial and Agricultural Biotechnologies, University of Verona, Verona, Italy. righetti@sci.univr.it
Castagna
Annalisa
A
Herbert
Ben
B
Candiano
Giovanni
G
D002458:Cell Fractionation / Q000379:methods; D003911:Dextrans / Q000737:chemistry; D004586:Electrophoresis / Q000295:instrumentation*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000295:instrumentation / Q000379:methods; D007525:Isoelectric Focusing / Q000295:instrumentation / Q000379:methods; D013058:Mass Spectrometry / Q000295:instrumentation; D008962:Models, Theoretical; D040901:Proteomics / Q000295:instrumentation / Q000379:methods*
D003911:Dextrans; C025614:sephadex