In cell biology, cell fractionation is the process used to separate cellular components while preserving individual functions of each component.[1] This is a method that was originally used to demonstrate the cellular location of various biochemical processes. Other uses of subcellular fractionation is to provide an enriched source of a protein for further purification, and facilitate the diagnosis of various disease states.[2]
Tissue is typically homogenized in a buffer solution that is isotonic to stop osmotic damage. Mechanisms for homogenization include grinding, mincing, chopping, pressure changes, osmotic shock, freeze-thawing, and ultrasound. The samples are then kept cold to prevent enzymatic damage. It is the formation of homogenous mass of cells (cell homogenate or cell suspension). It involves grinding of cells in a suitable medium in the presence of certain enzymes with correct pH, ionic composition, and temperature. For example, pectinase which digests middle lamella among plant cells.
Filtrationedit
This step may not be necessary depending on the source of the cells. Animal tissue however is likely to yield connective tissue which must be removed. Commonly, filtration is achieved either by pouring through gauze or with a suction filter and the relevant grade ceramic filter.
Purificationedit
Purification is achieved by differential centrifugation – the sequential increase in gravitational force results in the sequential separation of organelles according to their density.
^Alberts, B; Johnson, A (2002). "Fractionation of Cells". Molecular Biology of the Cell. 4th edition.
^Ninfa, Alexander J. (2010). Fundamental Laboratory Approaches for Biochemistry and Biotechnology. United States of America: John Wiley & Sons, INC. p. 209. ISBN978-0-470-08766-4.
January 01, 1970
cell, fractionation, cell, biology, cell, fractionation, process, used, separate, cellular, components, while, preserving, individual, functions, each, component, this, method, that, originally, used, demonstrate, cellular, location, various, biochemical, proc. In cell biology cell fractionation is the process used to separate cellular components while preserving individual functions of each component 1 This is a method that was originally used to demonstrate the cellular location of various biochemical processes Other uses of subcellular fractionation is to provide an enriched source of a protein for further purification and facilitate the diagnosis of various disease states 2 Contents 1 Homogenization 2 Filtration 3 Purification 4 See also 5 ReferencesHomogenization editTissue is typically homogenized in a buffer solution that is isotonic to stop osmotic damage Mechanisms for homogenization include grinding mincing chopping pressure changes osmotic shock freeze thawing and ultrasound The samples are then kept cold to prevent enzymatic damage It is the formation of homogenous mass of cells cell homogenate or cell suspension It involves grinding of cells in a suitable medium in the presence of certain enzymes with correct pH ionic composition and temperature For example pectinase which digests middle lamella among plant cells Filtration editThis step may not be necessary depending on the source of the cells Animal tissue however is likely to yield connective tissue which must be removed Commonly filtration is achieved either by pouring through gauze or with a suction filter and the relevant grade ceramic filter Purification editPurification is achieved by differential centrifugation the sequential increase in gravitational force results in the sequential separation of organelles according to their density See also editCell disruption Media for cell separation by density Percoll FicollReferences edit Alberts B Johnson A 2002 Fractionation of Cells Molecular Biology of the Cell 4th edition Ninfa Alexander J 2010 Fundamental Laboratory Approaches for Biochemistry and Biotechnology United States of America John Wiley amp Sons INC p 209 ISBN 978 0 470 08766 4 Retrieved from https en wikipedia org w index php title Cell fractionation amp oldid 1140871150, wikipedia, wiki, book, books, library,
