【摘要】 目的探讨双向凝胶电泳(2-DE)样品裂解液对蛋白质分离和双向凝胶电泳结果的影响。方法以MOLT-4细胞为例,采用3种溶解性能不同的裂解液提取细胞中的蛋白质组,并分别进行双向聚丙烯酰胺凝胶电泳。结果通过对2-D PAGE图谱的比较分析,发现采用8 mol/L Urea,2 mol/L Thiourea,4%CHAPS,1%NP-40,4%TritonX-100,65 mmol/L DTT,0.5 mmol/L PMSF,0.5%pharmalyte 3-10,能获得质量较高的2-DE图谱。结论采用高浓度脲、三去污裂解液有利于获得优质的蛋白质样品,从而提高蛋白质提取率和双向电泳分辨率等。更多还原

【Abstract】 Objective To investigate the effects of the sample lysis solution on protein separation and results about two-dimensional electrophoresis in two-dimensional gel electrophoresis.Methods Three different resolvable solutions were adopted to take out the proteins of MOLT-4 cells as the example,and then the proteins were separated by two dimensional polyacrylamide gel electrophoresis.Results By using 8 mol/L Urea,2 mol/L Thiourea,4 % CHAPS,1 % NP-40, 4 % TritonX-100,65 mmol/L DTT,0.5 mmol/L PMSF,0.5 % pharmalyte 3-10 much better quality map of 2-DE could be obtained.Conclusion Using lysis solution containing high concentration urea and three different detergents can obtain more quality samples,then increase the extraction rate of proteins and resolution about two-dimensional electrophoresis.更多还原