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  • The preparation of subcellular organelles from mouse liver in self-generated gradients of iodixanol.

The preparation of subcellular organelles from mouse liver in self-generated gradients of iodixanol.

Analytical biochemistry (1994-08-01)
J Graham, T Ford, D Rickwood
摘要

This paper reports the use of a new density gradient compound, Iodixanol, for the resolution of the major organelles from mouse liver. A major advantage of Iodixanol over other iodinated density gradient media is its ready ability to form self-generated gradients. Gradient-forming conditions have been modulated to provide optimal recoveries of Golgi 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. Iodixanol provided resolution superior to that of Nycodenz, notably of peroxisomes and mitochondria and the separation of lysosomes from endoplasmic reticulum. Because Iodixanol does not interfere significantly with marker enzyme activities, gradient fractions can be analyzed without removal of the gradient medium.

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Sigma-Aldrich
OptiPrep密度梯度培养基, used for cell and subcellular organelle isolation
Sigma-Aldrich
过氧化物酶体分离试剂盒, isolate peroxisomes from tissues and cells