Detergents and chaotropes for protein solubilization before two-dimensional electrophoresis.
Because of the outstanding ability of two-dimensional electrophoresis to separate complex mixtures of intact proteins, it would be advantageous to apply it to all types of proteins, including hydrophobic and membrane proteins. Unfortunately, poor solubility hampers the analysis of these molecules. As these problems arise mainly in the extraction and isoelectric focusing steps, the solution is to improve protein solubility under the conditions prevailing during isoelectric focusing. This chapter describes the use of chaotropes and novel detergents to enhance protein solubility during sample extraction and isoelectric focussing, and discusses the contribution of these compounds to improving proteomic analysis of membrane proteins.
Rabilloud
Thierry
T
Laboratoire, d'immunologie, INSERM, Grenoble, France.
D005591:Chemical Fractionation / Q000379:methods*; D003902:Detergents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional; D004912:Erythrocytes / Q000737:chemistry; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D007525:Isoelectric Focusing / Q000379:methods; D008565:Membrane Proteins / Q000737:chemistry* / Q000302:isolation & purification; D012995:Solubility; D013890:Thiourea / Q000737:chemistry*; D014508:Urea / Q000737:chemistry*
D003902:Detergents; D008565:Membrane Proteins; D014508:Urea; D013890:Thiourea