A simple procedure for the isolation of rat kidney lysosomes

Abstract

A simple procedure for the isolation of rat kidney lysosomes.BackgroundA procedure for the isolation of highly purified lysosomes from normal rat kidney is described.MethodsThe method depends on the swelling of mitochondria when the postnuclear supernatant fraction is incubated with 2mm Ca2+. The lysosomes can then be separated from the swollen mitochondria by Percoll density gradient centrifugation.ResultsThe 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.ConclusionWe believe that this procedure for isolating kidney lysosomes will be useful in the study of the mechanisms of specific modification, processing and catabolism of proteins