Abstract
In iodixanol, peroxisomes are the densest organelle in the light mitochondrial fraction and are therefore easily separated from the other components (lysosomes, mitochondria, etc.) in a preformed isosmotic continuous gradient. Because of the large difference in density between peroxisomes and the next densest organelle (mitochondria), a density barrier is effective. The resolution of the peroxisomes is far superior than that in sucrose and, unlike in Percoll®, there is no contamination from endoplasmic reticulum.
Highlights
Peroxisomes can be purified in iodixanol gradients in high yield (80–90%) with no detectable contamination from any other organelle[1,2,3]
In Nycodenz and metrizamide, mitochondria have a significantly higher density than in iodixanol because only the latter can provide an iso-osmotic medium at densities above ρ = 1.15–1.16 g/ml
The protocol below is for the liver from a young adult male rat, other tissues may require a different homogenization technique
Summary
Peroxisomes can be purified in iodixanol gradients in high yield (80–90%) with no detectable contamination from any other organelle[1,2,3]. This is a property unique to iodixanol because the densities of other organelles, that of mitochondria In Nycodenz and metrizamide, mitochondria have a significantly higher density Ρ = 1.165 g/ml) than in iodixanol because only the latter can provide an iso-osmotic medium at densities above ρ = 1.15–1.16 g/ml.
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