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A simple procedure for the isolation of rat kidney lysosomes.
- Source :
-
Kidney international [Kidney Int] 1998 Jul; Vol. 54 (1), pp. 275-8. - Publication Year :
- 1998
-
Abstract
- Background: A procedure for the isolation of highly purified lysosomes from normal rat kidney is described.<br />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.<br />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.<br />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.
- Subjects :
- Albumins analysis
Animals
Arylsulfatases metabolism
Catalase metabolism
Cathepsin D metabolism
Centrifugation
Colloids
Electron Transport Complex II
Glucose-6-Phosphatase metabolism
Lysosomes chemistry
Lysosomes enzymology
Male
Microscopy, Electron
Multienzyme Complexes metabolism
Oxidoreductases metabolism
Povidone
Rats
Rats, Wistar
Silicon Dioxide
Succinate Dehydrogenase metabolism
Cell Fractionation methods
Kidney cytology
Lysosomes ultrastructure
Subjects
Details
- Language :
- English
- ISSN :
- 0085-2538
- Volume :
- 54
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Kidney international
- Publication Type :
- Report
- Accession number :
- 9648089
- Full Text :
- https://doi.org/10.1046/j.1523-1755.1998.00958.x