| 肼化辣根过氧化物酶糖蛋白染色法 |
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| 引用本文： | 巨同忠,黄勇,陈惠黎.肼化辣根过氧化物酶糖蛋白染色法[J].复旦学报(医学版),1996(5). |
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| 作者姓名： | 巨同忠 黄勇 陈惠黎 |
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| 作者单位： | 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，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，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 |
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| 基金项目： | 国家自然科学基金!（3904000）资助项目 |
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| 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。
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| 关 键 词： | 辣根过氧化物酶 肼化物 糖蛋白 |
A METHOD FOR THE STAINING OF GLYCOPROTEIN BY HYDRAZIDE LINKED HORSERADISH PEROXIDASE |
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| Ju Tongzhong,Huang Yong,Chen Huili.A METHOD FOR THE STAINING OF GLYCOPROTEIN BY HYDRAZIDE LINKED HORSERADISH PEROXIDASE[J].Fudan University Journal of Medical Sciences,1996(5). |
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| Authors: | Ju Tongzhong Huang Yong Chen Huili |
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| 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. |
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| Keywords: | horseradish peroxidase hydrazide glycoprotein |
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