Isolation of peroxisomes from rat liver using sucrose and Percoll gradients.
Peroxisomes were isolated from the livers of both control and clofibrate-treated rats. Two procedures, one with a sucrose gradient, and a second with Percoll gradients, were utilized. The Percoll procedure allowed contamination of the isolated peroxisome fraction on protein basis, by lysosomes (8%), by mitochondria (5%) and by microsomes (2%). The peroxisome fraction isolated by the sucrose gradient showed no significant contamination with mitochondria, but the fraction contained 13% microsomes. In addition to established peroxisomal enzymes, the isolated peroxisomes also contained cytochrome b5, NADH-cytochrome c reductase and NADPH-isocitrate dehydrogenase. The peroxisomal membranes were also separated from the content, and they were found to have a relatively high phospholipid/protein ratio (0.55).
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002499:Centrifugation, Density Gradient / Q000379:methods; D002994:Clofibrate / Q000494:pharmacology; D003102:Colloids; D008099:Liver / Q000187:drug effects / Q000648:ultrastructure*; D008297:Male; D008830:Microbodies / Q000187:drug effects / Q000648:ultrastructure*; D008854:Microscopy, Electron; D009940:Organoids / Q000648:ultrastructure*; D011205:Povidone; D051381:Rats; D012822:Silicon Dioxide; D013395:Sucrose
D003102:Colloids; D013395:Sucrose; C016039:Percoll; D012822:Silicon Dioxide; D011205:Povidone; D002994:Clofibrate