High performance two-dimensional gel electrophoresis using a wetting agent Tergitol NP7.
Two-dimensional electrophoresis is an efficient method for the analysis of a broad range of complex protein samples. Current two-dimensional gel techniques are not suited for analysis of the small amount of proteins from tissue samples in the presence of high concentration of salts. Here we describe an improved two-dimensional gel electrophoresis procedure based on the use of a nonionic wetting agent, Tergitol NP7, in rehydration solution combined with the application of a linear potential sweep during isoelectrofocusing. This experimental approach yields a dramatic increase in the resolution and focusing of proteins visualized on two-dimensional gels. This technique is less time-consuming and laborious than the current techniques and can be used for a variety of two-dimensional electrophoresis applications, including proteome analysis.
Laoudj-Chenivesse
Dalila
D
Association Française contre les Myopathies, CNRS, Institut de Génétique Humaine, Montpellier, France. dlaoudj@igh.cnrs.fr
Marin
Philippe
P
Bennes
Rene
R
Tronel-Peyroz
Emmanuel
E
Leterrier
François
F
D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D007525:Isoelectric Focusing; D009132:Muscles / Q000737:chemistry; D011060:Poloxalene / Q000737:chemistry*; D020543:Proteome / Q000032:analysis; D012492:Salts / Q000737:chemistry; D013501:Surface-Active Agents / Q000737:chemistry*; D014020:Tissue Extracts / Q000032:analysis
D020543:Proteome; D012492:Salts; D013501:Surface-Active Agents; D014020:Tissue Extracts; D011060:Poloxalene