A simple procedure for the isolation of highly purified lysosomes from normal rat liver.
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.
D000818:Animals; D002118:Calcium; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D006867:Hydrolases / Q000032:analysis; D008099:Liver / Q000648:ultrastructure*; D008247:Lysosomes / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008854:Microscopy, Electron; D011205:Povidone; D051381:Rats; D011919:Rats, Inbred Strains; D012822:Silicon Dioxide
C016039:Percoll; D012822:Silicon Dioxide; D006867:Hydrolases; D011205:Povidone; D002118:Calcium