用¹²⁵I-阳离子化细胞色素c检测固定在未修饰和带正电荷的尼龙66上的蛋白聚糖和糖胺聚糖。

Detection by 125I-cationized cytochrome c of proteoglycans and glycosaminoglycans immobilized on unmodified and on positively charged nylon 66.

作者信息

Heimer R, Molinaro L, Sampson P M

机构信息

Department of Biochemistry, Thomas Jefferson University, Philadelphia, Pennsylvania 19107.

出版信息

Anal Biochem. 1987 Sep;165(2):448-55. doi: 10.1016/0003-2697(87)90295-8.

DOI:10.1016/0003-2697(87)90295-8

Abstract

We have examined the detection by a 125I-labeled basic protein, cationized cytochrome c, of selected proteoglycans (PGs) and standard preparations of glycosaminoglycans (GAGs) immobilized on Nylon 66 and also on positively charged Nylon 66. Immobilization on Nylon 66 appears to allow a relative freedom of interaction between PGs or GAGs and 125I-cationized cytochrome c, but a more restricted reaction was observed when PGs and GAGs were immobilized to positively charged Nylon 66. On this support PGs with large numbers of GAG side chains reacted well with 125I-cationized cytochrome c, but GAGs were minimally reactive. By taking advantage of some of the properties of large-pore agarose-acrylamide gels, rapid partial characterization of some PGs can be accomplished in the 10-ng range, and therefore at a sensitivity equal to PGs with internal biosynthetic labels.

摘要

我们检测了用¹²⁵I标记的碱性蛋白、阳离子化细胞色素c对固定在尼龙66以及带正电荷的尼龙66上的特定蛋白聚糖（PGs）和糖胺聚糖（GAGs）标准制剂的检测情况。固定在尼龙66上似乎能使PGs或GAGs与¹²⁵I - 阳离子化细胞色素c之间有相对自由的相互作用，但当PGs和GAGs固定在带正电荷的尼龙66上时，观察到的反应受到更多限制。在这种载体上，带有大量GAG侧链的PGs与¹²⁵I - 阳离子化细胞色素c反应良好，但GAGs的反应性极小。利用大孔琼脂糖 - 丙烯酰胺凝胶的一些特性，可在10纳克范围内对一些PGs进行快速部分表征，因此灵敏度与带有内部生物合成标记的PGs相当。