Yamada H, Hayashi H, Natori Y
J Biochem. 1984 Apr;95(4):1155-60. doi: 10.1093/oxfordjournals.jbchem.a134704.
A procedure for the isolation of highly purified lysosomes from normal rat liver is described. The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 1 mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation. The lysosomal fraction obtained by our method was enriched more than 120-fold in terms of the marker enzymes with a yield of 25%. The electron microscopic examination and the measurement of the activities of marker enzymes for various subcellular organelles indicated that our lysosomal preparation was essentially free from contamination by other organelles.
本文描述了一种从正常大鼠肝脏中分离高纯度溶酶体的方法。该方法基于当核后上清液与1 mM Ca2+孵育时线粒体的肿胀。然后可以通过Percoll密度梯度离心将溶酶体与肿胀的线粒体分离。用我们的方法获得的溶酶体组分,就标记酶而言,富集了120多倍,产率为25%。电子显微镜检查和各种亚细胞器标记酶活性的测量表明,我们制备的溶酶体基本上没有其他细胞器的污染。