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Methods Mol Biol. 2008;424:35-42. doi: 10.1007/978-1-60327-064-9_3.

Sample solublization buffers for two-dimensional electrophoresis.

Author information

1
Technical University of Munich, Munich, Germany.

Abstract

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.

PMID:
18369850
DOI:
10.1007/978-1-60327-064-9_3
[Indexed for MEDLINE]

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