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Hybridoma Cultures for Production of Antibodies

Purification and Specificity of Antibodies

  • Protocol
Protocols for Neural Cell Culture

Part of the book series: Springer Protocols Handbooks ((SPH))

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Abstract

During the process of characterizing the structure and function of novel antigens, it is usually necessary to create new monoclonal or polyclonal antibody reagents. However, once validated, these antibodies can be put to a multitude of experimental uses, such as detecting and quantitating antigens in cell and tissue extracts or biological fluids, purifying proteins for structural analyses, studying protein-protein interactions, and monitoring cellular differentiation. Although many well-characterized monoclonal antibodies (mAbs) to antigens of general interest are commercially available, mAbs to specialty antigens may need to be individually purified from hybridomas obtained from academic sources, or from not-for-profit cell repositories, such as the American Type Culture Collection (http://www.atcc.org). This chapter describes protocols for producing, purifying, and verifying murine mAbs from pre-existing hybridomas.

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Further Reading

  • Akerstrom, B., Brodin, T., Reis, K., and Bjorck, L. (1985), Protein G: a powerful tool for binding and detection of monoclonal and polyclonal antibodies. J. Immunol. 135, 2589–2592.

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  • Kawamoto, T., Sato, J. D., McClure, D. B., and Sato, G. H. (1986), Serum-free medium for the growth of NS-1 mouse myeloma cells and the isolation of NS-1 hybridomas. Methods Enzymol. 121, 266–277.

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Author information

Authors and Affiliations

  1. Division of Cell, Molecular, and Developmental Biology, American Type Culture Collection, Manassas, VA

    J. Denry Sato

Authors
  1. J. Denry Sato
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Editor information

Editors and Affiliations

  1. Department of Anatomy and Cell Biology, School of Medicine, University of Saskatchewan, Saskatoon, Saskatchewan, Canada

    Sergey Fedoroff  & Arleen Richardson  & 

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© 2001 Humana Press Inc., Totowa, NJ

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Sato, J.D. (2001). Hybridoma Cultures for Production of Antibodies. In: Fedoroff, S., Richardson, A. (eds) Protocols for Neural Cell Culture. Springer Protocols Handbooks. Humana Press. https://doi.org/10.1385/1-59259-207-4:317

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  • DOI: https://doi.org/10.1385/1-59259-207-4:317

  • Publisher Name: Humana Press

  • Print ISBN: 978-0-89603-902-5

  • Online ISBN: 978-1-59259-207-4

  • eBook Packages: Springer Book Archive

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