Thornton D J, Howard M, Devine P L, Sheehan J K
Division of Biochemistry, School of Biological Sciences, University of Manchester, United Kingdom.
Anal Biochem. 1995 May 1;227(1):162-7. doi: 10.1006/abio.1995.1266.
Mucins (density 1.35-1.50 g/ml) prepared from human cervical, gastric, respiratory, and salivary secretions were reduced and alkylated to generate reduced mucin subunits. These subunits were separated into different populations analytically by using agarose gel electrophoresis and preparatively by using anion-exchange chromatography. Both techniques separate the molecules primarily on the basis of their inherent charge. A procedure has been developed for the efficient deglycosylation of small amounts of these reduced mucin subunits immobilized on polyvinylidene difluoride. The combination of these procedures has allowed the isolation and identification of glycoforms of a specific mucin gene product in a respiratory mucous secretion.
从人宫颈、胃、呼吸道和唾液分泌物中制备的粘蛋白（密度1.35 - 1.50克/毫升）经还原和烷基化处理，生成还原型粘蛋白亚基。这些亚基通过琼脂糖凝胶电泳进行分析分离，并通过阴离子交换色谱进行制备分离。这两种技术主要根据分子的固有电荷来分离分子。已开发出一种程序，用于对固定在聚偏二氟乙烯上的少量这些还原型粘蛋白亚基进行高效去糖基化。这些程序的组合使得能够在呼吸道粘液分泌物中分离和鉴定特定粘蛋白基因产物的糖型。
