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Resolubilization of TCA precipitated plant proteins for 2-D electrophoresis
Erhua Zhang1, Xiaoping Chen, Xuanqiang Liang
1Crops Research Institute, Guangdong Academy of Agricultural Science, Wushan, Guangzhou, P. R. China.
Abstract:
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.
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