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  • Title: A simple procedure for the isolation of rat kidney lysosomes.
    Author: Kawashima A, Sato A, Kawashima M, Nitta K, Yumura W, Sugino N, Nihei H, Natori Y.
    Journal: Kidney Int; 1998 Jul; 54(1):275-8. PubMed ID: 9648089.
    Abstract:
    BACKGROUND: A procedure for the isolation of highly purified lysosomes from normal rat kidney is described. METHODS: The method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 2 mM Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation. RESULTS: The lysosomal fraction obtained by our method was enriched more than 30-fold in terms of marker enzymes with a yield of about 11%. 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. CONCLUSION: We believe that this procedure for isolating kidney lysosome will be useful in the study of the mechanisms of specific modification, processing and catabolism of proteins.
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