【摘要】 对双向电泳方法进行了改进,包括对样品的制备、双向电泳参数的选择等关键步骤进行优化.实验发现,采用乙酸铵-甲醇沉淀拟南芥全细胞蛋白质,同时结合高伏时、长时间的等电聚焦可以获得高分辨率、重复性好的蛋白质双向电泳图谱.银染后,经Phoretix 2D v2004软件分析可分辨出1 000以上蛋白点.更多还原

【Abstract】 An optimized protocol was reported with special attention focused on the whole-cell protein analyses based on two-dimensional gel electrophoresis(2-DE) and Arabidopsis as material.The optimization included sample preparation,electrophoresis conditions,gel staining,protein spots resolution and reproducibility.The results indicate that in combination with high-voltage and long-running time in IEF,the precipitation of(proteins) with ammonium acetate/methanol method will give rise to satisfactory 2-DE image.The maximal(detectable) protein spots were estimated to be more than 1 000 under silver staining.This protocol may be(applicable) to other plant materials for 2-DE studies.更多还原