墓头回提取物作用白血病K562细胞蛋白双向电泳方法的建立

o optimize K562 cell sample preparation, optimal parameters of two-dimensional electrophoresis were established to improve resolution and reproducibility which is used to investigate the mechanism between diversifolious patrinia root and K562 cell. Standard lysis buffer were used to preparing K562 cell samples. Two different preparation methods, TCA/ Acetone precipitation and acetone precipitation, were applied to K562 cell samples and the acetone precipitation method was proved to be the better one considering the resheults obtained from two-dimensional gel electrophoresis (2-DE). Acetone precipitation not only decreased protein decomposition，but also increased protein solubility，therefore obtaining the two dimensional electrophoresis spectrum with higher resolution and better reproducibility. Besides, the focus voltage for gel electrophoresis was optimized on the traditional focus conditions and a two-dimensional electrophoresis system with a clear background and good protein separation was successfully established for leukemia K562 cells. The optimized focusing condition made horizontal and vertical tail improved obviously, and a high-resolution two-dimensional electrophoresis map with clearer protein spots and more complete separation obtained easily. Acetone precipitation on the basic of standard lysis buffer was more suitable for preparing the sample of K562 cell to operate two-dimensional electrophoresis. In addition, the longer desalinization time in isoelectric focusing (IEF) can separate whole protein effectively and the Following tests forcefully proves the wide utility of the optimized methods for the tumor tissues .