Purification of a crude mitochondrial fraction by density-gradient centrifugation.
Most mitochondria prepared by differential centrifugation are contaminated to some extent by lysosomes, peroxisomes, tubular Golgi membranes, and small amounts of endoplasmic reticulum. Density gradient centrifugation using a variety of density media--sucrose, Percoll, Nycodenz, Iodixanol--is described here and is used to prepare purer fractions of mitochondria. The resulting gradient can be analyzed for three marker enzymes: succinate dehydrogenase (mitochondria), b-galactosidase (lysosomes), and catalase (peroxisomes).
Graham
J M
JM
Liverpool John Moores University, Liverpool, United Kingdom.
D000818:Animals; D015415:Biomarkers; D001921:Brain; D002374:Catalase / Q000032:analysis; D002458:Cell Fractionation / Q000379:methods*; D002499:Centrifugation, Density Gradient / Q000379:methods*; D008247:Lysosomes / Q000201:enzymology; D008297:Male; D008928:Mitochondria / Q000201:enzymology; D008930:Mitochondria, Liver / Q000201:enzymology; D020675:Peroxisomes / Q000201:enzymology; D011205:Povidone; D051381:Rats; D017207:Rats, Sprague-Dawley; D012822:Silicon Dioxide; D013385:Succinate Dehydrogenase / Q000032:analysis; D014283:Triiodobenzoic Acids; D001616:beta-Galactosidase / Q000032:analysis
D015415:Biomarkers; D014283:Triiodobenzoic Acids; C016039:Percoll; D012822:Silicon Dioxide; D002374:Catalase; D013385:Succinate Dehydrogenase; D001616:beta-Galactosidase; D011205:Povidone; C044834:iodixanol