摘要
双向电泳技术是目前最常用的能够从植物组织中分离出上千种蛋白的技术．因此，需对蛋白样品制备过程、等电聚焦、第二相电泳、染色等进行分析探讨，结果表明，样品制备需根据样品的成分选择不同的裂解液成分（如：CHAPS，TritonX-100和DTT等）；胶条越长要求上样量越大（考染上样量约为银染上样的5倍），等电聚焦电压越大，聚焦过程也越长；SDS-PAGE胶体积分数为10％～15％，常用12％；第二相电泳需选择恒定电流，且先以10mA／胶，跑0．5～1．0h，再以大电流30mA／胶至第二相电泳完成．
2-DE is the most commonly used technology to separate hundreds of proteins from the plant tissues.We analyzed and studied the processes of sample preparation,IFE,second-phase electrophoresis,gel-staining.The result showed that the sample preparation due to different elements (CHAPS,Triton X-100,DTT etc)should be selected according to different pyrolysis liquid elements.Longer of gel requires more loading amount of sample (5 times loading amount of sample in G250-stainning more than silver-stainning).Higher isoelectric focusing voltage the longer is the process of focusing.The volume fraction of SDS-PAGE is 10%-15%,commonly used 12%.polyacrylamide gel.The second -phase electrophoresis should select constant current and firstly 10 mA /gel should be set to run 0.5-1.0 h then 30 mA /gel till second-phase electrophoresis was finished.
出处
《昆明学院学报》
2015年第3期69-71,86,共4页
Journal of Kunming University
基金
云南省教育厅资助项目(2014Y391)
关键词
双向电泳
蛋白质
电泳图谱
等电聚焦
聚丙烯酰胺凝胶
two-dimensional electrophoresis
protein
electrophoretogram
isoelectric focusing
polyacrylamide gel
