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Protein Blotting and Detection pp 293–298Cite as

Sequential Use of Immunoblots for Characterization of Autoantibody Specificities

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 536))

Summary

Sera of patients with systemic autoimmune diseases frequently contain autoantibodies to nuclear autoantigens. Immunoblotting of recombinant and native autoantigens is a commonly used technique for the identification and characterization of autoantibody specificities. Here, we describe an easy procedure that facilitates the comparison of antibody specificities by reusing the same immunoblot at least three times in order to detect an abundantly expressed autoantigen in total cellular extracts.

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References

  1. Tan, A.N. (1989) Antinuclear antibodies: diagnostic markers for autoimmune diseases and probes for cell biology. Adv Immunol. 44, 93–151.

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  2. Laemmli, U.K. (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680–685.

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Acknowledgements

Special thanks for excellent technical support to K. Joanne and Tim Gross (Oklahoma Medical Research Foundation, OK, USA).

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Authors and Affiliations

  1. Institute Immunology, MTZ, Medical Faculty, Carl Gustav Carus University TU Dresden, Fiedlerstrasse 42, D-01307, Dresden, Germany

    Holger Bartsch & Michael Bachmann

Authors
  1. Holger Bartsch
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  2. Michael Bachmann
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Corresponding author

Correspondence to Michael Bachmann .

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Editors and Affiliations

  1. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    Biji T. Kurien

  2. Oklahoma Medical Research Foundation, NE. 13th Street 825, Oklahoma City, 73104, U.S.A.

    R. Hal Scofield

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© 2009 Humana Press, a part of Springer Science+Business Media, LLC

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Bartsch, H., Bachmann, M. (2009). Sequential Use of Immunoblots for Characterization of Autoantibody Specificities. In: Kurien, B., Scofield, R. (eds) Protein Blotting and Detection. Methods in Molecular Biology, vol 536. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-59745-542-8_31

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  • DOI: https://doi.org/10.1007/978-1-59745-542-8_31

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  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-1-934115-73-2

  • Online ISBN: 978-1-59745-542-8

  • eBook Packages: Springer Protocols

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