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目的:初步建立并优化用于蛋白质组分析的双向电泳技术,提高其分辨率及重复性。方法:以HepG2细胞为例,对以固相pH梯度为第一向的双向电泳的关键因素与环节,如样品处理、电泳参数和凝胶浓度进行一系列的优化。结果与结论:本研究采用增溶、排异的裂解法处理样品,选择适中的样品量,用预制固相pH梯度——IPG胶条(pH=3~10)第进行第一向等电聚焦,并选用12%~14%的梯度胶进行第二向SDS分离,可获得质量较好的双向电泳图谱
OBJECTIVE: To initially establish and optimize two-dimensional electrophoresis for proteomic analysis to improve its resolution and reproducibility. Methods: Taking HepG2 cells as an example, a series of optimization of key factors and links of two-dimensional electrophoresis, such as sample processing, electrophoresis parameters and gel concentration, were carried out. RESULTS AND CONCLUSION: In this study, the samples were treated by solubilization and rejection method. The samples were selected moderately, and the first-dimension isoelectric focusing was carried out by pre-solid phase pH gradient - IPG strips (pH = 3-10) , And the selection of 12% to 14% of the gradient gel for the second SDS separation, get better quality two-dimensional electrophoresis