Abstract
The ability to generate antibodies that recognize a given protein relies on that protein’s “antigenicity”, i.e., its ability to appear foreign to the host immune system. Thus, proteins that are highly conserved among species are often poor antigens when used for immunization (1). This lack of immunoreactivity is not owing to the inherent nature of the protein; rather, it is because of the mechanism of tolerance, which exists to minimize autoimmune disease. Immunological tolerance occurs in an organism to prevent the development of a destructive immune response to self-proteins. It operates by eliminating or inactivating immune cells that bear receptors that recognize and respond to autologous protein antigens (2). Thus, it is very difficult to generate an immune response in mice to either murine (i.e., self) antigens or to highly conserved proteins (1).
This is a preview of subscription content, log in via an institution.
Buying options
Tax calculation will be finalised at checkout
Purchases are for personal use only
Learn about institutional subscriptionsSpringer Nature is developing a new tool to find and evaluate Protocols. Learn more
References
Goding, J. W. (1983) Monoclonal Antibodies: Principles and Practice. In: Production and application of monoclonal antibodies in cell biology, biochemistry, and immunology. Aca-demic Press, London, UK, pp. 5–55.
Burnet, F. M. (1959) The Clonal Selection Theory of Acquired Immunity. Vanderbilt Uni-versity Press, Nashville, TN, USA.
Thomas, K. R. and Capecchi, M. R. (1987) Site-directed mutagenesis by gene targeting in mouse embryo-derived stem cells. Cell. 51, 503–512.
Claesson, M. H., Endel, B., Ulrik, J., Pedersen, L. O., Skov, S., and Buus, S. (1994). Anti-bodies directed against monomorphic and evolutionary conserved self epitopes may be generated in ’knock-out’ mice. Development of monoclonal antibodies directed against monomorphic MHC class I determinants. Scand. J. Immunol. 40(2), 257–264.
Castrop, J., Verbeek, S., Hofhuis, F., and Clevers, H. (1995) Circumvention of tolerance for the nuclear T cell protein TCF-1 by immunization of TCF-1 knock-out mice. Immunobiology 193, 281–287.
Declerck, P.J., Carmeliet, P., Verstreken, M., De Cock, F., and Collen, D. (1995). Genera-tion of monoclonal antibodies against autologous proteins in gene-inactivated mice. J. Biol. Chem. 270, 8397–8400.
Warren, S.T., and Nelson, D. L. (1994) Advances in molecular analysis of fragile X syn-drome. JAMA 271, 536–542.
Jin, P. and Warren, S. T. (2000) Understanding the molecular basis of fragile X syndrome. Hum. Mol. Gen. 9, 901–908.
Ashley, C. T., Sutcliffe, J. S., Kunst, C. B., Leiner, H. A., Eichler, E. E., Nelson, D. L., and Warren, S. T. (1993) Human and murine FMR-1: alternative splicing and translational initiation downstream of the CGG repeat. Nat. Genet. 4, 244–251.
Price, D. K., Zhang, F., Ashley, C. T., and Warren, S. T. (1996) The chicken FMR1 gene is highly conserved with a CCT 5′-untranslated repeat and encodes an RNA-binding protein. Genomics. 31, 3–12.
Siomi, M., Siomi, H., Sauer, W. H., Srinivasan, S., Nussbaum, R. L., and Dreyfuss, G. (1995) FXR1, an autosomal homolog of the fragile X mental retardation gene. EMBO J. 14, 2401–2408.
Devys, D., Lutz, Y., Rouyer, N., Bellocq, J.-P., and Mandel, J.-L. (1993) The FMR-1 protein is cytoplasmic, most abundant in neurons, and appears normal in carriers of the fragile X premutation. Nat. Genet. 4, 335–340.
Harlow, E. and Lane, D. (1988) Antibodies: A laboratory manual. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA.
Coligan, J. E., Kruisbeek, A. M., Margulies, D. M., Shevach, E. M., and Strober, W. (1999) Current Protocols in Immunology. John Wiley and Sons, Inc., New York, NY, USA.
Roitt, I. (1988) Essential Immunology, 6th ed., Blackwell Scientific Publication, Oxford, UK.
Anonymous. (1994). Fmr1 knockout mice: a model to study fragile X mental retardation. Cell. 78(1), 23–33.
Ceman, S., Brown, V., and Warren., S. T. (1999) Isolation of an FMRP-associated messen-ger ribonucleoprotein particle and identification of nucleolin and the fragile X-related pro-teins as components of the complex. Mol. Cell. Biol. 19(12), 7925–7932.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2003 Humana Press Inc.
About this protocol
Cite this protocol
Ceman, S., Zhang, F., Johnson, T., Warren, S.T. (2003). Development and Characterization of Antibodies that Immunoprecipitate the FMR1 Protein. In: Potter, N.T. (eds) Neurogenetics. Methods in Molecular Biology™, vol 217. Springer, Totowa, NJ. https://doi.org/10.1385/1-59259-330-5:345
Download citation
DOI: https://doi.org/10.1385/1-59259-330-5:345
Publisher Name: Springer, Totowa, NJ
Print ISBN: 978-0-89603-990-2
Online ISBN: 978-1-59259-330-9
eBook Packages: Springer Protocols