蛋白质电泳非样品条带来源分析  

Assay the source of non-sample bands in polyacrylamide gel electrophoresis

作　　者：周海涛[1] 曾华书[1] 马伟钦[2] 侯红斌[1] 陈润莉[1] 张勇[1] 李康[2] 

机构地区：[1]深圳市福田区疾病预防控制中心,广东深圳518040 [2]广东药学院,广东广州510310

出　　处：《癌变．畸变．突变》2013年第3期232-235,共4页Carcinogenesis,Teratogenesis & Mutagenesis

摘　　要：目的:分析十二烷基硫酸钠-聚丙烯酰胺凝胶电泳中非样品条带的来源,并探讨其消除及控制方法。方法:在每个凝胶梳孔只加全成分上样缓冲液或缺少某些成分的试液,不加蛋白样品直接电泳,观察非样本条带出现情况,再分别对上样缓冲液中还原剂、甘油成分设置成不同浓度梯度进行电泳分析,观察条带表现。另外观察不同的上样梳预处理对条带表现的影响,明确非样品条带来源,找到去除方法。最后,进一步通过副溶血弧菌蛋白电泳结果验证非样品条带控制方法的有效性。结果:在未对上样梳进行特别处理的情况下,上样试液(全成分缓冲液或缺少某些成分的试液)中只要有还原剂和甘油就会出现非样品条带,但条带强弱与还原剂浓度、甘油含量无明显剂量关系。用8mol/L尿素处理上样梳可使非样品条带明显减弱甚至消失。结论:非样品条带源于上样梳蛋白质污染,用尿素等离液剂或强力去污剂处理上样梳可有效减少或消除非样品条带,以消除非样品条带对检验结果的影响。OBJECTIVE： To explore the sources and control methods of non-sample bands in the protein SDS-polyacrylamide gel electrophoresis. METHODS：The sample loading buffer,and its different component parts without protein sample were directly added to the gel and electrophoresed to identify the non-sample bands. Impact on non-sample bands of gradient concentrations of reducing agent and glycerol in the buffer were also assessed. The effective methods for removing the non-sample bands were further verified by pretreatment comb for preparation gel. Finally,method for reducing non-sample bands was tested by SDS-PAGE electrophoresis of protein of Vibrio parahaemolyticus. RESULTS：Non-sample bands appeared as long as the reducing agent and glycerol were both present in the buffer solution with comb pretreated by ordinary commercially available detergents,but the intensity of the bands showed no significant dose relationship with the concentration of reducing agent or glycerol. Treating the comb with 8 mol/L urea liquid,the non-sample bands showed weakening or even disappeared. CONCLUSION：Non-sample bands were caused by pollution of the comb. Treating the comb with a strong protein dissolving agent,such as 8 mol/L urea may be useful to reduce as eliminate non-sample bands.

关 键 词：十二烷基硫酸钠-聚丙烯酰胺凝胶电泳 非样品条带 上样缓冲液 污染 

分 类 号：TQ937[化学工程]