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, inc...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 ug 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.展开更多
基金funded by the National Natural Science Foundation of China (Grant No.30860141)
文摘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 ug 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.
