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文摘
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比较了聚乙二醇修饰蛋白体系的SDS-聚丙烯酰胺凝胶电泳(SDS-PAGE)银染、考染、碘化钡染色3种染色方法;提出和比较了银染-碘化钡复染和考染-碘化钡复染2种复染方法.结果表明,银染-碘化钡复染的凝胶中,未修饰蛋白条带消失,PEG修饰蛋白条带保留,游离PEG条带显色;而考染-碘化钡复染的凝胶中,未修饰蛋白、修饰蛋白和游离的PEG条带可同时显色.两种复染方法中,PEG组分的检测限均达到了0.01μg.因此,对PEG修饰蛋白体系的SDS-PAGE可先用考染或银染后再用碘化钡复染,便可在同一块凝胶上先后或同时观察到未修饰蛋白、修饰蛋白和游离PEG的情况,简化了实验操作,方便了实验结果的比较分析. |
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其他语种文摘
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Three kinds of staining methods for PEGylated protein system in SDS-PAGE were compared, and two kinds of restaining methods were developed and compared. The gels were stained with coomassie brilliant blue staining, and silver staining, and then restained by BaI_2 staining. PEGylated protein and PEG were observed when the gels were restained by BaI_2; after silver staining, while the native protein band disappeared. On the other hand, all of the native protein, PEGylated protein and PEG were observed when it was restained by BaI_2, after coomassie brilliant blue staining, so all of the components of the PEGylated protein system could be seen on one gel in succession or at the same time. The detection limit of PEGylated protein is 0.01μg in each restaing. It could reduce half of the work and the cost of the gels, avoid the operating error on different gels, and make the observation of the results more convenient. |
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来源
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化学通报
,2009,72(5):459-462 【核心库】
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关键词
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PEG修饰
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电泳
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SDS-PAGE
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碘化钡染色
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复染
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地址
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中国科学院过程工程研究所, 生化工程国家重点实验室, 北京, 100080
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语种
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中文 |
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文献类型
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简报 |
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ISSN
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0441-3776 |
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学科
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化学 |
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基金
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国家自然科学基金青年基金
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国家高技术研究发展计划(863计划)
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北京市自然科学基金
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文献收藏号
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CSCD:3530089
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