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Identification of theĀ Antigen Content of Electroimmunoprecipitates

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Immunoproteomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2024))

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Abstract

Polyclonal antibodies including purified antibody fractions and animal or human antisera may react with unknown antigens or antigens other than their main specificity in reactions that are best visualized by gel electroimmunoprecipitation methods (e.g., when analyzing complex antigen mixtures). The great advantage of gel immunoprecipitation approaches is that each immunoprecipitate contains antigen in a pure form and that the precipitate is separated by position, shape, and size from other precipitates in the complex patterns of crossed immunoelectrophoresis. The identification of the antigen content of such immunoprecipitates is important but challenging because of the very stable, high-affinity complex formation leading to precipitation in the gels. Here, we present detailed step-by-step recipes for identifying the antigen content of electroimmunoprecipitates.

This article is dedicated to Niels H. H. Heegaard, who passed away suddenly during the preparation of this manuscript.

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Beyer, N.H., Heegaard, N.H.H. (2019). Identification of theĀ Antigen Content of Electroimmunoprecipitates. In: Fulton, K., Twine, S. (eds) Immunoproteomics. Methods in Molecular Biology, vol 2024. Humana, New York, NY. https://doi.org/10.1007/978-1-4939-9597-4_7

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  • DOI: https://doi.org/10.1007/978-1-4939-9597-4_7

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-4939-9596-7

  • Online ISBN: 978-1-4939-9597-4

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