title: Evaluation of protocols used in 2-d electrophoresis for proteome analysis of young rice caryopsis.
abstract: In order to obtain a high-resolution electrophorogram of rice young panicle proteome, we evaluated various protocols commonly used in two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) of proteins, including gel staining protocol, pH range of immobilized pH gradient (IPG) strips and sample loading quantity. Results showed that a silver staining protocol using sensitized solution containing glacial acetic acid, sodium acetate and sodium thiosulfate (reported by Heukeshoven and Dernick in 1988) and a Coomassie Brilliant Blue staining method using solution containing G-250, ammonium sulfate and phosphoric acid (reported by Pink et al in 2010) demonstrated the superior staining effect. In addition, we also showed that higher resolution was achieved when IPG gel strip with pH range of 5-8 was used, compared to that with pH range of 4-7. Finally, the optimal loading quantity was determined as 130 μg using the 17 cm-long nonlinear IPG strip with pH 5-8 in combination with the silver nitrate staining protocol. The evaluated results would be helpful in proteome analysis of young rice caryopsis.
authors.affiliation: MOE Key Laboratory of Crop Physiological Ecology and Genetic Breeding, Jiangxi Agricultural University, Nanchang 330045, China.
authors.forename: Jiang-Lin
authors.initials: JL
authors.lastname: Liao
authors.forename: Ying-Jin
authors.initials: YJ
authors.lastname: Huang
mesh_terms: D015180:Electrophoresis, Gel, Two-Dimensional / Q000379:methods; D004591:Electrophoresis, Polyacrylamide Gel; D006863:Hydrogen-Ion Concentration; D012275:Oryza / Q000378:metabolism*; D010756:Phosphoric Acids; D011506:Proteins / Q000378:metabolism; D020543:Proteome / Q000737:chemistry / Q000378:metabolism*; D040901:Proteomics / Q000379:methods*; D012394:Rosaniline Dyes; D016622:Silver Staining
chemical_list: D010756:Phosphoric Acids; D011506:Proteins; D020543:Proteome; D012394:Rosaniline Dyes; C030242:phosphoric acid; C004692:coomassie Brilliant Blue