Sample solublization buffers for two-dimensional electrophoresis.
Before two-dimensional electrophoresis (2-DE), proteins of the sample must be denatured, reduced, disaggregated, and solubilized. Sample solubilization is usually carried out in a buffer containing chaotropes (typically 9.5 M urea, or 5-8 M urea and 2 M thiourea), 2-4% nonionic and/or zwitterionic detergent(s), reducing agent(s), carrier ampholytes and, depending on the type of sample, protease inhibitors. In this chapter, the major constituents of sample solubilization/lysis buffers will be briefly reviewed, some general sample preparation guidelines will be given, and the most common protein solubilization cocktails will be described.
Weiss
Walter
W
Technical University of Munich, Munich, Germany.
Görg
Angelika
A
D002021:Buffers; D003902:Detergents / Q000737:chemistry*; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D017408:Guidelines as Topic; D011489:Protein Denaturation; D013048:Specimen Handling / Q000379:methods*; D013890:Thiourea / Q000737:chemistry*; D014508:Urea / Q000737:chemistry*
D002021:Buffers; D003902:Detergents; D014508:Urea; D013890:Thiourea