title: Resolubilization of TCA precipitated plant proteins for 2-D electrophoresis.
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.
authors.affiliation: Crops Research Institute, Guangdong Academy of Agricultural Science, Wushan, Guangzhou, P. R. China.
authors.forename: Erhua
authors.initials: E
authors.lastname: Zhang
authors.forename: Xiaoping
authors.initials: X
authors.lastname: Chen
authors.forename: Xuanqiang
authors.initials: X
authors.lastname: Liang
mesh_terms: D000478:Alkylation; D010367:Arachis / Q000737:chemistry; D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods*; D019791:Guanidine; D010720:Phosphines / Q000737:chemistry; D018515:Plant Leaves / Q000737:chemistry; D010940:Plant Proteins / Q000737:chemistry* / Q000302:isolation & purification / Q000378:metabolism; D011725:Pyridines / Q000737:chemistry; D015203:Reproducibility of Results; D016622:Silver Staining; D012995:Solubility; D019032:Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; D014238:Trichloroacetic Acid / Q000737:chemistry*
chemical_list: D010720:Phosphines; D010940:Plant Proteins; D011725:Pyridines; C043255:tri-n-butylphosphine; D014238:Trichloroacetic Acid; C030953:2-vinylpyridine; D019791:Guanidine