当前位置: 首页 > 文章 > 适于SDSPAGE分析的高粱叶片蛋白质提取方法 河南农业科学 2013,42 (5) 38-40
Position: Home > Articles > Protein Extraction Methods of Sorghum Leaf Suitable for SDS-PAGE Analysis Journal of Henan Agricultural Sciences 2013,42 (5) 38-40

适于SDSPAGE分析的高粱叶片蛋白质提取方法

作  者:
许克静;陈云;雷珍珍;程海丽;陈磊;乐超银
单  位:
三峡大学化学与生命科学学院
关键词:
高粱;SDS-聚丙烯酰胺凝胶电泳;蛋白质提取;样品制备
摘  要:
为了探索适于高粱叶片蛋白质SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)分析的样品制备方法,对TCA/丙酮沉淀后的蛋白质分别用裂解液Ⅰ(尿素+硫脲+CHAPS+DTT)和裂解液Ⅱ(尿素+DTT)2种裂解液进行裂解,用改良的Bradford法进行定量分析,并进行SDS-PAGE分离检测。结果表明,裂解液Ⅰ所得蛋白质质量浓度(2.538g/L)高于裂解液Ⅱ所得蛋白质质量浓度(1.905g/L),且两者呈极显著差异(P<0.05),但裂解液Ⅰ所得蛋白质电泳条带出现1条杂带;而裂解液Ⅱ所得蛋白质能满足上样要求且电泳条带清晰无杂带。因此,TCA/丙酮沉淀并用裂解液Ⅱ进行裂解的方法适用于高粱叶片的SDS-PAGE分析。
译  名:
Protein Extraction Methods of Sorghum Leaf Suitable for SDS-PAGE Analysis
作  者:
XU Ke-jing,CHEN Yun,LEI Zhen-zhen,CHENG Hai-li,CHEN Lei,YUE Chao-yin(College of Chemistry and Life Science,China Three Gorges University,Yichang 443000,China)
关键词:
sorghum;SDS-PAGE;protein extraction;sample preparation
摘  要:
In order to develop a sample preparation method suitable for SDS-PAGE analysis of sorghum leaf protein,the protein(after TCA/aceton precipitation) was resolubilized with lysisbufferⅠ(urea,thiourea,CHAPS and DTT) and lysis buffer Ⅱ(urea,DTT),respectively.And then the protein was quantified with modified Bradford assay,and was seperated by SDS-PAGE.The results showed that the concentration of protein(2.538 g/L) got from lysis bufferⅠwas higher than that from bufferⅡ(1.905 g/L),the difference being significant(P<0.05).But anunexpected band of protein electrophoresis was appeared from lysis buffer Ⅰ;whereas,enough protein was got from lysis buffer Ⅱ,and the bands were sharp and clear with no extra band.So the method of resolubilizing TCA/aceton precipitated protein with lysis bufferⅡ was suitable for SDS-PAGE analysis of sorghum leaf.

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