Ugozzoli M, Chiu A
Division of Neurosciences, Beckman Research Institute of the City of Hope, Duarte, CA 91010.
Biotechniques. 1992 Feb;12(2):187-8, 190.
Several components of the extracellular matrix in the molecular weight range of 220 kDa to 150 kDa were purified by preparative electrophoresis on 2.5% Pro-Sieve agarose gels. These high molecular weight glycoproteins, separated under reducing conditions, were recovered in solution by extraction of individual agarose gel slices and analyzed on sodium dodecyl sulfate polyacrylamide gels and Western blots. This simple method permitted the separation and recovery of the laminin B chains (220 kDa and 205 kDa) and entactin (150 kDa) and may prove useful for the purification of other high molecular weight species.
通过在2.5%的Pro-Sieve琼脂糖凝胶上进行制备电泳,纯化了分子量在220 kDa至150 kDa范围内的细胞外基质的几种成分。这些在还原条件下分离的高分子量糖蛋白,通过提取单个琼脂糖凝胶切片在溶液中回收,并在十二烷基硫酸钠聚丙烯酰胺凝胶和蛋白质免疫印迹上进行分析。这种简单的方法可以分离和回收层粘连蛋白B链(220 kDa和205 kDa)和巢蛋白(150 kDa),可能对纯化其他高分子量物质有用。