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Development and use of antiproteasome monoclonal antibodies

Klavs B Hendil1

  • 1Institute of Molecular Biology and Physiology, University of Copenhagen, DK-2100 Copenhagen, Denmark.

Methods in Enzymology
|November 9, 2005
PubMed

This chapter describes the production of hybridomas and screening of monoclonal antibodies for use in enzyme-linked immunosorbent assay (ELISA), immunoblotting, and immunoprecipitation of components of the ubiquitin-proteasome pathway. Purification of antibodies and their use in affinity chromatography are also described.

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Hybridoma Technology

Hybridoma technology is used for the large-scale production of monoclonal antibodies. Monoclonal antibodies bind to only a single antigenic determinant or epitope. Such antibodies are used in research, diagnostics, and disease therapy. The hybridoma technology established in 1975 by Georges Köhler and Cesar Milstein was awarded the Nobel Prize in Medicine in 1984 for revolutionizing research and therapy.
Hybridoma Selection
Commonly used fusion techniques — electroporation, polyethylene glycol...