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. 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
This Protocol Article describes the use of a preformed continuous gradient in a fixed-angle rotor
TheScientificWorldJOURNAL (2002) 2, 1536-1539 mitochondria have a higher density in Nycodenz or metrizamide than in iodixanol, the resolution of these two organelles is much easier in iodixanol gradients[5,6,7]
Summary
Peroxisomes can be purified in iodixanol gradients in high yield (80–90%) with no detectable contamination from any other organelle. This Protocol Article describes the use of a preformed continuous gradient in a fixed-angle rotor. Peroxisomes are the densest of the major subcellular organelles (ρ = 1.18–1.20 g/ml) present in the light mitochondrial fraction from mammalian tissues and cells.
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