BIOCYTIN HYDRAZIDE - A SELECTIVE LABEL FOR SIALIC ACIDS, GALACTOSE, AND OTHER SUGARS IN GLYCOCONJUGATES USING AVIDIN BIOTIN TECHNOLOGY
BIOCYTIN HYDRAZIDE - A SELECTIVE LABEL FOR SIALIC ACIDS, GALACTOSE, AND OTHER SUGARS IN GLYCOCONJUGATES USING AVIDIN BIOTIN TECHNOLOGY
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DOI：
10.1016/0003-2697(88)90631-8
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发表时间：
1988-05-01
影响因子：
2.9
通讯作者：
WILCHEK, M
中科院分区：
文献类型：
--
作者：
BAYER, EA;BENHUR, H;WILCHEK, M
生物胞素酰肼（BCHZ）是一种新型的水溶性长链生物素酰肼，可用于糖缀合物的选择性非放射性检测。开发了用于在印迹上标记糖缀合物的程序。该方法涉及糖缀合物的化学（高碘酸盐诱导）或酶促（通过半乳糖氧化酶）氧化，然后用生物胞素酰肼标记所得醛基，然后与基于抗生物素蛋白的酶探针相互作用。 由于含生物素的试剂是一种相对较小的带电分子，因此可以在完整细胞和膜制备物以及印迹样品上进行初步标记步骤。在印迹，标记模式是相似的高碘酸盐和半乳糖氧化酶诱导的生物素化程序。与此相反，高碘酸盐诱导标记的红细胞膜或细胞（前印迹）产生了改变标记模式。组合酶诱导的膜或细胞的生物素化导致的模式类似于观察到的直接染色的印迹。使用人红细胞膜上的半乳糖氧化酶，该程序足够灵敏，可以选择性地标记带3乳糖胺糖蛋白。
Biocytin hydrazide (BCHZ), a new, water-soluble, long-chained, biotin-containing hydrazide, was synthesized and used for the selective nonradioactive detection of glycoconjugates. Procedures were developed for labeling glycoconjugates on blots. The method involves either chemical (periodate-induced) or enzymatic (via galactose oxidase) oxidation of glycoconjugates, the resultant aldehyde groups are then labeled with biocytin hydrazide, followed by interaction with an avidin-based enzyme probe. Since the biotin-containing reagent is a relatively small, charged molecule, the primary labeling step may be carried out on intact cells and on membrane preparations as well as on blotted samples. On blots, the labeling pattern was similar for both periodate- and galactose oxidase-induced biotinylation procedures. In contrast, periodate-induced labeling of either erythrocyte membranes or cells (prior to blotting) produced an altered labeling pattern. Combined enzyme-induced biotinylation of membranes or cells resulted in a pattern similar to that observed for the direct staining of blots. Using galactose oxidase on human erythrocyte membranes, the procedure was sensitive enough to selectively label the Band 3 lactosaminoglycoprotein.