Zhu Q, Bhavanandan V P
Department of Biochemistry and Molecular Biology, Milton S. Hershey Medical Center, Pennsylvania State University, Hershey 17033, USA.
Glycoconj J. 1995 Oct;12(5):639-44. doi: 10.1007/BF00731259.
Alkaline borohydride treatment is widely used for the release of carbohydrate moieties from O-glycosylated glycoproteins and mucins. We have adapted this procedure to micro quantities of glycoproteins blotted on membranes. After electrophoresis and transfer to nitrocellulose, nylon or polyvinylidene difluoride membrane, alkaline borohydride treatment was done directly on glycoprotein containing areas of membrane which were cut out with the aid of guide strips stained with Coomassie Blue or lectin-digoxigenin. In combination with standard saccharide fractionation techniques, this procedure can be used to characterize the oligosaccharides of mucins or mucin-type glycoproteins that are separated by gel electrophoresis from crude sources. Using this approach we have characterized the saccharides derived from the two species of epitectin, a malignancy-associated mucin type glycoprotein, isolated from metabolically labelled H.Ep2 cells.
碱性硼氢化物处理被广泛用于从O-糖基化糖蛋白和粘蛋白中释放碳水化合物部分。我们已将此方法应用于印迹在膜上的微量糖蛋白。在电泳并转移至硝酸纤维素膜、尼龙膜或聚偏二氟乙烯膜后，借助考马斯亮蓝或凝集素-地高辛配基染色的引导条带，直接对膜上含有糖蛋白的区域进行碱性硼氢化物处理。结合标准的糖类分级技术，该方法可用于表征通过凝胶电泳从粗提物中分离出的粘蛋白或粘蛋白型糖蛋白的寡糖。使用这种方法，我们已对源自两种表位蛋白（一种与恶性肿瘤相关的粘蛋白型糖蛋白）的糖类进行了表征，该表位蛋白是从经代谢标记的H.Ep2细胞中分离得到的。
