von Boxberg Y, Wütz R, Schwarz U
Max-Planck-Institut für Entwicklungsbiologie, Abteilung Biochemie, Tübingen, Federal Republic of Germany.
Eur J Biochem. 1990 Jun 20;190(2):249-56. doi: 10.1111/j.1432-1033.1990.tb15569.x.
We describe a method for the selective labelling, isolation and electrophoretic analysis of cell-surface molecules and extracellular matrix components. Intact tissues are reacted with activated esters of biotin and the labelled surface molecules identified on Western blots with horseradish-peroxidase-coupled or 35S-labelled streptavidin. Alternatively, the biotinylated proteins can be purified from tissue homogenates by affinity chromatography on an avidin-agarose column. Evidence is presented to show that this method is indeed specific for membrane and matrix components. Its practical application to embryonic neural tissues is demonstrated.
我们描述了一种用于细胞表面分子和细胞外基质成分的选择性标记、分离及电泳分析的方法。完整组织与生物素的活化酯反应,然后在蛋白质印迹法中用辣根过氧化物酶偶联的或35S标记的抗生物素蛋白鉴定标记的表面分子。或者,可以通过在抗生物素蛋白-琼脂糖柱上进行亲和层析从组织匀浆中纯化生物素化的蛋白质。有证据表明该方法确实对膜成分和基质成分具有特异性。还展示了其在胚胎神经组织中的实际应用。