Preparation of Extracts From Animal Tissues

  • J. Mark Skehel
Part of the Methods in Molecular Biology book series (MIMB, volume 244)


The initial procedure in the isolation of an protein, a protein complex, or a subcellular organelle is the preparation of an extract that contains the required component in a soluble form. Indeed, when undertaking a proteomic study, the production of a suitable cellular extract is essential. Further isolation of subcellular fractions depends on the ability to rupture the animal tissues in such a manner that the organelle or macromolecule of interest can be purified in a high yield, free from contaminants and in an active form. The homogenization technique employed should, therefore, stress the cells sufficiently enough to cause the surface plasma membrane to rupture, thus releasing the cytosol; however, it should not cause extensive damage to the subcellular structures, organelles, and membrane vesicles. The extraction of proteins from animal tissues is relatively straightforward, as animal cells are enclosed only by a surface plasma membrane (also referred to as the limiting membrane or cell envelope) that is only weakly held by the cytoskeleton. They are relatively fragile compared to the rigid cell walls of many bacteria and all plants and are thus susceptible to shear forces. Animal tissues can be crudely divided into soft muscle (e.g., liver and kidney) or hard muscle (e.g., skeletal and cardiac). Reasonably gentle mechanical forces such as those produced by liquid shear may disrupt the soft tissues, whereas the hard tissues require strong mechanical shear forces provided by blenders and mincers. The homogenate produced by these disruptive methods is then centrifuged in order to remove the remaining cell debris.


Animal Tissue Integral Membrane Protein Surface Plasma Membrane Rigid Cell Wall Mince Tissue 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


  1. 1.
    Claude, A. (1946) Fractionation of mammalian liver cells by differential centrifugation: II. Experimental procedures and results. J. Exp. Med. 84, 61–89.CrossRefGoogle Scholar
  2. 2.
    Rabilloud, T. (1995) A practical guide to membrane protein purification. Electrophoresis 16(3), 462–471.CrossRefGoogle Scholar
  3. 3.
    Arigita, C., Jiskoot, W., Graaf, M. R., and Kersten, G. F. A. (2001) Outer membrane protein purification. Methods Mol. Med. 66, 61–79.PubMedGoogle Scholar
  4. 4.
    Smith, A. L. (1967) Preparation, properties and conditions for assay of mitochondria: slaughterhouse material, small scale. Methods Enzymol. 10, 81–86.CrossRefGoogle Scholar
  5. 5.
    Tyler, D. D. and Gonze, J. (1967) The preparation of heart mitochondria from laboratory animals. Methods Enzymol. 10, 75–77.CrossRefGoogle Scholar
  6. 6.
    Dignam, J. D. (1990) Preparation of extracts from higher eukaryotes. Methods Enzymol. 182, 194–203.PubMedCrossRefGoogle Scholar
  7. 7.
    Völkl, A. and Fahimi, H. D. (1985) Isolation and characterization of peroxisomes from the liver of normal untreated rats. Eur. J. Biochem. 149, 257–265.PubMedCrossRefGoogle Scholar
  8. 8.
    Ernster, L. and Nordenbrand, K. (1967) Skeletal muscle mitochondria. Methods Enzymol. 10, 86–94.CrossRefGoogle Scholar
  9. 9.
    Scarpa, A., Vallieres, J., Sloane, B., and Somlyo, A. P. (1979) Smooth muscle mitochondria. Methods Enzymol. 55, 60–65.PubMedCrossRefGoogle Scholar
  10. 10.
    Blobel, G. and Potter, V. R. (1966) Nuclei from rat liver: isolation method that combines purity with high yield. Science 154, 1662–1665.PubMedCrossRefGoogle Scholar
  11. 11.
    Nedergaard, J. and Cannon, B. (1979) Overview—preparation and properties of mitochrondria from different sources Methods Enzymol. 55, 3–28.PubMedCrossRefGoogle Scholar
  12. 12.
    Chappell, J. B. and Perry, S. V. (1954) Biochemical and osmotic properties of skeletal muscle mitochondria. Nature 173, 1094–1095.PubMedCrossRefGoogle Scholar

Copyright information

© Humana Press Inc., Totowa, NJ 2004

Authors and Affiliations

  • J. Mark Skehel
    • 1
  1. 1.Genomic and Proteomic Sciences, Medicines Research CenterGlaxoSmithKline PharmaceuticalsStevenageUK

Personalised recommendations