A rapid method for the isolation of peroxisomes from rat liver.
A preparative method for the isolation of peroxisomes from the liver of normal, untreated rats is described. The peroxisome-enriched "light mitochondrial" fraction is layered on a 30% Nycodenz (5-[N-2,3-dihydroxypropylacetamido]-2,4,6-triiodo-N,N'-bis[2, 3-dihydroxypropyl]isophthalamide) solution containing 1 mM tetrasodium EDTA and then centrifuged in an angular rotor for 1 h at 130,000gavg. Peroxisomes are sedimented to the bottom leaving other organelles at the top of the tube. On the basis of morphological and biochemical studies, it is found that the peroxisomes (marker-enzymes catalase and urate oxidase) obtained in this method are not contaminated with lysosomes (marker-enzyme acid phosphatase) and contained very few mitochondria (marker-enzyme succinate-cytochrome c reductase) and microsomal vesicles (marker-enzyme glucose-6-phosphatase).
D000217:Acyltransferases / Q000032:analysis; D000818:Animals; D002374:Catalase / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient; D008099:Liver / Q000201:enzymology / Q000648:ultrastructure*; D008297:Male; D008830:Microbodies / Q000201:enzymology / Q000648:ultrastructure*; D051381:Rats; D014503:Urate Oxidase / Q000032:analysis
D002374:Catalase; D014503:Urate Oxidase; D000217:Acyltransferases; C020051:glycerone-phosphate O-acyltransferase