以便获得米饭幼仔圆锥花序 proteome 的高分辨率的 electrophorogram,我们评估了通常使用在的各种各样的协议二维(2D ) 蛋白质的 polyacrylamide 胶化电气泳动(页) 包括染色协议的胶化,使不能调动的 pH 坡度(IPG ) 的 pH 范围脱衣并且样品装载数量。结果证明染色协议使用的银敏化包含冰川的醋酸,钠醋酸盐和钠 thiosulfate 的答案(在 1988 由 Heukeshoven 和 Dernick 报导了)并且染色方法使用答案包含的 Coomassie 灿烂的蓝色 G-250 ,铵硫酸盐和磷的酸(在 2010 由粉红色的等报导了)表明了优异染色效果。另外,当有 5-8 的 pH 范围的 IPG 胶化长带被使用时,我们也与 4-7 的 pH 范围证明更高的分辨率被完成,与那相比。最后,最佳的装载数量作为与染色协议的银硝酸盐在联合与 pH 5-8 使用 17 厘米长的非线性的 IPG 长带的 130 g 被决定。评估结果将在年轻米饭颖果的 proteome 分析是有用的。
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.