Ligler F S, Robinson G B
Biochim Biophys Acta. 1977 Aug 1;468(3):27-40.
A method is described for the isolation of basement membranes from rabbit renal cortex in which the detergent N-lauroyl sarcosine is used as the disruptive agent. The isolated membranes have been compared with membranes prepared using ultrasonication and they were comparable both in terms of purity and gross chemical composition. Glomerular and tubular basement membranes were isolated by first separating glomeruli from tubules by density gradient centrifugation followed by detergent treatment of the separated tissues. The detergent method has the advantage that the basement membranes retained their native structure to a large degree, whereas sonicated membranes were severely fragmented. Collagen fibres were a significant contaminant in both preparations and were revealed more clearly by negative staining than by examination of thin sections. Studies with the detergent-treated membrane revealed that a few proteins, which seemed to be membrane components, were extracted with 1 M NaCl and that these proteins were lost from the basement membranes during sonication used in the conventional isolation procedure.
本文描述了一种从兔肾皮质中分离基底膜的方法，该方法使用去污剂N-月桂酰肌氨酸作为破坏剂。将分离得到的膜与通过超声处理制备的膜进行了比较，在纯度和总体化学成分方面二者具有可比性。通过密度梯度离心首先将肾小球与肾小管分离，然后对分离的组织进行去污剂处理，从而分离出肾小球和肾小管基底膜。去污剂法的优点是基底膜在很大程度上保留了其天然结构，而超声处理的膜则严重破碎。在两种制备物中胶原纤维都是一种重要的污染物，通过负染色比通过薄片检查更清楚地显示出来。对经去污剂处理的膜的研究表明，一些似乎是膜成分的蛋白质可被1M NaCl提取出来，并且在传统分离程序中使用的超声处理过程中，这些蛋白质从基底膜中丢失。
