Enzyme-linked immunosorbent assay (ELISA)-based quantification and identification of in vitro enzyme-catalyzed glycosphingolipid synthesis and degradation products with carbohydrate sequence-specific monoclonal antibodies.
Enzyme-linked immunosorbent assay (ELISA)-based quantification and identification of in vitro enzyme-catalyzed glycosphingolipid synthesis and degradation products with carbohydrate sequence-specific monoclonal antibodies.
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使用碳水化合物序列特异性单克隆抗体，基于酶联免疫吸附测定 (ELISA) 对体外酶催化鞘糖脂合成和降解产物进行定量和鉴定。
DOI：
10.1016/0003-2697(88)90530-1
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发表时间：
1988
影响因子：
2.9
通讯作者：
Macher,BA
中科院分区：
文献类型：
--
作者：
Stults,CL;Wilbur,BJ;Macher,BA
建立了一种新的检测糖基转移酶和糖基水解酶活性的方法。反应产物的鉴定和定量是用酶联免疫吸附试验（ELISA）使用碳水化合物序列特异性单克隆抗体同时完成的。用β-半乳糖基转移酶和α-半乳糖苷酶反应来说明该方法的显著特点。这些包括简单的产品鉴定和定量，不需要洗涤剂，消耗少量试剂，不使用放射性同位素。此外，可以用该方法测量底物消失或产物形成。用该方法研究了酶的Km、Vmax、二价阳离子需要量和最适pH等特性。
A new method has been developed to monitor glycosyltransferase and glycosylhydrolase activities. Reaction product identification and quantification are accomplished simultaneously with an enzyme-linked immunosorbent assay (ELISA) using carbohydrate sequence-specific monoclonal antibodies. β-Galactosyltransferase and α-galactosidase reactions were used to illustrate the salient features of the method. These include simple product identification and quantification, no detergent requirement, consumption of small amounts of reagents, and no use of radioisotopes. Furthermore, it is possible to measure substrate disappearance or product formation with this method. enzyme characteristics such as Km, Vmax, divalent cation requirement, and pH optimum were investigated with this new method.