Loten E G, Redshaw-Loten J C
Anal Biochem. 1986 Apr;154(1):183-5. doi: 10.1016/0003-2697(86)90512-9.
Existing procedures for the preparation of rat liver plasma membranes are time consuming and generally produce low yields. A method is described in which a rat liver homogenate low-speed pellet is fractionated on a self-forming Percoll gradient. Plasma membranes can be removed from the gradient in a high yield along with much of the DNA in the liver homogenate. A second Percoll step performed in the presence of a low concentration of calcium ions separates the DNA from the plasma membranes. The final membrane fraction has high specific activities of marker enzymes with little contamination with microsomal, mitochondrial, Golgi, or lysosomal markers.
现有的制备大鼠肝细胞膜的方法耗时且产率普遍较低。本文描述了一种方法,即对大鼠肝匀浆低速沉淀在自形成的Percoll梯度上进行分级分离。细胞膜可与肝匀浆中的大部分DNA一起从梯度中以高产率分离出来。在低浓度钙离子存在下进行的第二步Percoll操作可将DNA与细胞膜分离。最终的膜组分具有标记酶的高比活性,几乎没有微粒体、线粒体、高尔基体或溶酶体标记物的污染。