Improved recovery of rat liver fractions enriched in lysosomes by specific alteration of the sedimentation properties of mitochondria.

摘要: A method for the preparation of lysosomes from rat liver is presented. The procedure requires only standard equipment and completed within less than 3 h. Homogenization differential centrifugation were performed at pH 7.4 in isotonic potassium phosphate-buffered sucrose medium. addition phosphate, concentration used (10 mM), accelerated sedimentation rate mitochondria without altering that resulting decrease mitochondrial contamination final pellet. Further purification was achieved by isopycnic 45% Percoll an angle rotor. Lysosomal fractions representing 51.5% original population recovered over a density range 1.09 to 1.15 g/ml. most purified fraction (37-fold purified) contained 25.3% lysosomal beta-N-acetylglucosaminidase, 0.9% monoamine oxidase 0.6% peroxisomal urate activities. It practically devoid endoplasmic reticulum contamination.