Abstract
Identification of proteins separated by gel electrophoresis or isoelectric-focusing is often compounded by the small pore size of the gel, which limits penetration by macromolecular probes. Overcoming this problem can be achieved by blotting the proteins onto an adsorbent porous membrane (usually nitrocellulose or diazo-tized paper), which gives a mirror image of the gel (1). A variety of reagents can be incubated with the membrane specifically to detect and analyze the protein of interest. Antibodies are widely used as detecting reagents, and the procedure is sometimes called Western blotting. However, protein blotting or immunoblotting is the most descriptive.
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Towbin, H., Staehelin, T., and Gordon, J. (1979) Electrophoretic transfer of proteins from polyacrylamide gels to nitrocellulose sheets: procedure and some applications. Proc. Natl. Acad. Sci. USA 76, 4350–4354.
Johnstone, A. and Thorpe, R. (1996) Immunochemistry in Practice, 3rd ed. Black-well Scientific, Oxford, UK.
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© 2009 Humana Press, a part of Springer Science+Business Media, LLC
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Page, M., Thorpe, R. (2009). Protein Blotting by Electroblotting. In: Walker, J.M. (eds) The Protein Protocols Handbook. Springer Protocols Handbooks. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-198-7_58
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DOI: https://doi.org/10.1007/978-1-59745-198-7_58
Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-60327-474-6
Online ISBN: 978-1-59745-198-7
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