Purification of peroxisomes from livers of normal and clofibrate-treated mice.
A method for the isolation of peroxisomes from livers of normal and clofibrate-treated mice is described. The method utilizes glutaraldehyde to stabilize peroxisomal membranes, and isopycnic centrifugation of a light mitochondrial fraction through a linear metrizamide gradient to achieve optimal resolution from other organelles. On the basis of the biochemical and morphological data, the peroxisomal preparations are indicated as of high purity: contamination by mitochondria, lysosomes, and plasma membranes is negligible, and the level of contaminating microsomes is around 5% for normal peroxisomes and 8% for peroxisomes from clofibrate-treated mice. Peroxisomal membranes prepared by carbonate extraction contain two major polypeptides of approximately 70,000 Da, and show 2 and 8% contamination by microsomal membrane protein for the preparations from normal and clofibrate-treated mice, respectively.
D000818:Animals; D002458:Cell Fractionation / Q000379:methods; D002994:Clofibrate / Q000494:pharmacology*; D005260:Female; D005976:Glutaral; D008099:Liver / Q000201:enzymology / Q000378:metabolism / Q000648:ultrastructure*; D008565:Membrane Proteins / Q000378:metabolism; D051379:Mice; D008830:Microbodies / Q000201:enzymology / Q000378:metabolism* / Q000648:ultrastructure; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism
D008565:Membrane Proteins; D002994:Clofibrate; D005976:Glutaral