昆明学院学报
昆明學院學報
곤명학원학보
JOURNAL OF KUNMING UNIVERSITY
2015年
3期
69-71,86
,共4页
程霞%窦玉敏%唐萍%苏源%邓纲
程霞%竇玉敏%唐萍%囌源%鄧綱
정하%두옥민%당평%소원%산강
双向电泳%蛋白质%电泳图谱%等电聚焦%聚丙烯酰胺凝胶
雙嚮電泳%蛋白質%電泳圖譜%等電聚焦%聚丙烯酰胺凝膠
쌍향전영%단백질%전영도보%등전취초%취병희선알응효
two-dimensional electrophoresis%protein%electrophoretogram%isoelectric focusing%polyacrylamide gel
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,Triton X-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10 mA /胶,跑0.5~1.0 h,再以大电流30 mA /胶至第二相电泳完成.
雙嚮電泳技術是目前最常用的能夠從植物組織中分離齣上韆種蛋白的技術.因此,需對蛋白樣品製備過程、等電聚焦、第二相電泳、染色等進行分析探討,結果錶明,樣品製備需根據樣品的成分選擇不同的裂解液成分(如:CHAPS,Triton X-100和 DTT 等);膠條越長要求上樣量越大(攷染上樣量約為銀染上樣的5倍),等電聚焦電壓越大,聚焦過程也越長;SDS-PAGE 膠體積分數為10%~15%,常用12%;第二相電泳需選擇恆定電流,且先以10 mA /膠,跑0.5~1.0 h,再以大電流30 mA /膠至第二相電泳完成.
쌍향전영기술시목전최상용적능구종식물조직중분리출상천충단백적기술.인차,수대단백양품제비과정、등전취초、제이상전영、염색등진행분석탐토,결과표명,양품제비수근거양품적성분선택불동적렬해액성분(여:CHAPS,Triton X-100화 DTT 등);효조월장요구상양량월대(고염상양량약위은염상양적5배),등전취초전압월대,취초과정야월장;SDS-PAGE 효체적분수위10%~15%,상용12%;제이상전영수선택항정전류,차선이10 mA /효,포0.5~1.0 h,재이대전류30 mA /효지제이상전영완성.
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.