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肼化辣根过氧化物酶糖蛋白染色法

A METHOD FOR THE STAINING OF GLYCOPROTEIN BY HYDRAZIDE LINKED HORSERADISH PEROXIDASE

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【作者】 巨同忠黄勇陈惠黎

【Author】 Ju Tongzhong, Huang Yong, Chen Huili (Key Laboratory of Glycoconjugate Resaerch, Department of Biochemistry, School of Basic Medical Sciences, Shanghai Medical University, Shanghai 200032)

【机构】 卫生部糖复合物重点实验室上海医科大学基础医学院生物化学教研室!200032

【摘要】 探索一种简便灵敏的糖蛋白半定量法。在 pH 5.0,碳二亚胺(EDC)催化下,辣根过氧化物酶(HRP)与己二酰二肼反应生成肼化的HRP(HRP-HZ),固相于硝酸纤维素膜上的糖蛋白(纤连蛋白或转铁蛋白)经过碘酸氧化其糖链中的糖基后生成的醛基可与HRP—HZ缩合,用HRP的发包底物显色即可检出精蛋白的存在与否。结果:糖蛋白中,10 ng的糖即可被检出。结论:与其他糖定量方法相比,该方法灵敏度高,方法简便,能够普遍应用。

【Abstract】 PURPOSE To estallish a simple sensitive method for the semiquantitative detection of glycoprotein. METHODS horseradish peroxidase (HRP) was reacted with adipic acid dihydrazide to form hydrazide linked HRP(HRP-HZ) at PH 5.0 under the catalyses of l-ethyl-3-(3’- dimethylaminopropyl)-carboimide(EDC). After the glyan of the glycoproteins (fibronectin or transferrin) immobilized on nitro-cellulose membranes were oxidized by sodium periodate (NaIO4), they can be conjugated with HRP-HZ and detected after the staining with HRP substrate. RESULTS Only 10ng of glycan can be clearly detected. CONCLUSIONS Compared with other quantitive methods for the determination of glycoprotein, this method is more sensitive and convenient to be widely used.

【关键词】 辣根过氧化物酶肼化物糖蛋白
【Key words】 horseradish peroxidasehydrazideglycoprotein
【基金】 国家自然科学基金!(3904000)资助项目
  • 【分类号】R341
  • 【被引频次】1
  • 【下载频次】131
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