Isolation of subcellular fractions
1997; Oxford University Press; Linguagem: Inglês
10.1093/oso/9780199634958.003.0002
AutoresRichard H. Hinton, Barbara M. Mullock,
Tópico(s)Lipid metabolism and biosynthesis
ResumoAbstract The aim of subcellular fractionation is to separate cell organelles with as little damage as possible. The first thing to remember is that it will never be possible to separate organelles completely undamaged. In the living cell, most of the cell organelles are attached to cytoskeletal elements and are surrounded by cytosol which is a 20% solution of protein in which most of the water is bound to the hydration sphere of the proteins. Breakage of the linkage to the cytoskeleton and alterations in the environment will thus inevitably result in alterations to the cell organelles and these alterations will undoubtedly be exacerbated by damage occurring during homogenization, as discussed in Chapter 1, and damage occurring during the course of separation, as discussed in this and subsequent chapters. Thus it will never be possible to separate cell organelles in a completely natural state and, in fact, there is never a single best way to fractionate a tissue. For example, the method used to determine the distribution of some material between cell organelles may differ markedly from the method used to isolate a particular organelle with the minimum of damage. The first aim of this chapter will be to discuss the general principles of both analytical and preparative methods for cell fractionation.
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