Preparation of Biotinylated Lectins and Application in Microtiter Plate Assays and Western Blotting
Preparation of Biotinylated Lectins and Application in Microtiter Plate Assays and Western Blotting
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生物素化凝集素的制备及其在微量滴定板测定和蛋白质印迹中的应用
DOI：
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发表时间：
1996
期刊：
影响因子：
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通讯作者：
A. M. Wu
中科院分区：
文献类型：
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作者：
E. Lisowska;M. Duk;A. M. Wu
凝集素通过氨基与生物胺基己酸N-羟基琥珀酰亚胺酯的生物素化提供了完全保留的碳水化合物结合活性的试剂，用酶标记的ExtrAvidin可以高灵敏度地检测到这些活性。在亲和素介导的检测中，生物素与生物素酰肼通过高碘酸盐氧化的碳水化合物残基进行生物素化是不太敏感的试剂；然而，后者可用于鉴定凝集素的糖基化状态。本文介绍了生物素/亲和素介导的凝集素微量平板法。由于生物素-亲和素相互作用的高亲和力，该方法能够比抗体介导的酶联免疫吸附试验(ELISA)更准确地测定结合到配体涂层平板上的凝集素。还描述了生物素化凝集素在检测印迹糖蛋白方面的应用，并用几个例子进行了说明。
The biotinylation of lectins via amino groups with biotinamidocaproate N-hydroxysuccinimide ester provided reagents with fully preserved carbohydrate-binding activity which were detectable with high sensitivity using enzyme-conjugated ExtrAvidin. Lectins biotinylated with biotin hydrazide via periodate-oxidized carbohydrate residues were less sensitive reagents in avidin-mediated assays; however, the latter method may serve for identification of the lectin glycosylation status. The biotin/avidin-mediated lectin microtiter plate assay is described. Due to the high affinity of biotin-avidin interaction, this assay enables a more accurate determination of the lectin bound to the ligand-coated plate than an antibody-mediated, enzyme-linked immunosorbent assay (ELISA). The application of biotinylated lectins for detection of glycoproteins on the blot is also described and illustrated with several examples.
人血型糖蛋白 A 在野生型和糖基化缺陷型中国仓鼠卵巢细胞中的表达。
DOI：
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发表时间：
1991
期刊：
The Journal of biological chemistry
影响因子：
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作者：
Remaley,AT;Ugorski,M;Wu,N;Litzky,L;Burger,SR;Moore,JS;Fukuda,M;Spitalnik,SL
通讯作者：
Spitalnik,SL