Human lysosomes can be purified from diploid skin fibroblasts by free-flow electrophoresis.
- 1 October 1980
- journal article
- research article
- Published by Proceedings of the National Academy of Sciences in Proceedings of the National Academy of Sciences
- Vol. 77 (10) , 6139-6143
- https://doi.org/10.1073/pnas.77.10.6139
Abstract
Human diploid fibroblasts underwent lysis when resuspended in isotonic sucrose. After preparation of a granular fraction by differential centrifugation, lysosomes were almost completely purified by free-flow electrophoresis. EM of the final fraction of fibroblast lysosomes (about 25-fold purified) showed mostly secondary lysosomes having an appearance identical to those in intact cells. There was no indication that the purification steps were selected for any lysosomal subpopulation. The lysosomal nature of the isolated organelles was confirmed by cytochemical staining for acid .beta.-glycerophosphatase. As judged by a 92% structure-linked latency of .beta.-N-acetylglucosaminidase in the final fraction, the structural integrity of the isolated lysosomes was unaffected by the purification procedure.Keywords
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