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Resolubilization of TCA precipitated plant proteins for 2‐D electrophoresis
Author(s) -
Zhang Erhua,
Chen Xiaoping,
Liang Xuanqiang
Publication year - 2011
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/elps.201000557
Subject(s) - solubility , guanidine , chemistry , silver stain , chromatography , electrophoresis , solubilization , hydrochloride , alkylation , gel electrophoresis , biochemistry , organic chemistry , biology , microbiology and biotechnology , catalysis
In this short communication, a novel protocol for resolubilization of TCA‐precipitated plant proteins for 2‐DE is described. Guanidine hydrochloride (Gdn‐HCl) is used as an intermediate for protein solubilization in which proteins are reduced and alkylated with tributylphosphane (TBP) and 2‐vinylpyridine (2‐VP). The blocking of –SH groups at Cys residues can greatly improve the solubility of TCA‐precipitated proteins and obtain more high‐quality protein spots in the 2‐DE gel. This protocol is compatible with silver stain and MS identification.