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The isolation of nuclear, cytosolic and mitochondrial fractions of a certain purity from mammalian tissues and cells has
attracted great interest due to their advantages in the study and characterization of different cellular proteins and organelles.
These subcellular fractions are commercially available from individual livers (inter-individual variation can be studied) or a pool
of livers (representative of the average enzyme activities of the population). They can be stored and remain stable at -80°C for
many years and can be thawed easily. They are very easy to handle for several types of incubations and they are suitable for
full automation. All of these advantages make them very useful for metabolism studies in the early stage of drug development
including the screening program.
Subcellular fractionation utilizes one or more of the properties of each compartment, such as buoyant density, surface charge
density, size, and shape, and is mainly based on differential centrifugation in a high viscosity media. Gel filtration, affinity
chromatography, electrophoresis or selective density-shift perturbation can also be used. Variations in the conditions of the
available protocols are dependent on the organelle, tissue or cell type and equipment used.
Fractionation by centrifugation