THE PREPARATION OF SUBCELLULAR ORGANELLES FROM MOUSE-LIVER IN SELF-GENERATED GRADIENTS OF IODIXANOL

This paper reports the use of a new density gradient compound, Iodixan ol, for the resolution of the major organelles from mouse liver. A maj or advantage of Iodixanol over other iodinated density gradient media is its ready ability to form self-generated gradients. Gradient-formin g conditions have been modulated to provide optimal recoveries of Golg i membranes, lysosomes, mitochondria, and peroxisomes. The organelles were isolated in high yield (80-90% of gradient input) and high purity . Nycodenz and Iodixanol were compared using preformed gradients. Iodi xanol provided resolution superior to that of Nycodenz, notably of per oxisomes and mitochondria and the separation of lysosomes from endopla smic reticulum. Because Iodixanol does not interfere significantly wit h marker enzyme activities, gradient fractions can be analyzed without removal of the gradient medium. (C) 1994 Academic Press, Inc.