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Antibody Specificity Profiling Using Protein Microarrays

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Epitope Mapping Protocols

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1785))

Abstract

Antibodies are the most widely used reagent for isolation and detection of specific proteins. However, using antibodies that are not highly specific in these studies can generate inaccurate and misleading data. Protein microarrays offer a platform by which antibody cross-reactivity against a broad range of cellular antigens can be simultaneously and quantitatively profiled. This protocol describes in detail the process of array pretreatment, antibody binding, washing, scanning and quantitative analysis of antibody specificity.

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Acknowledgments

This work was supported by NIH grant U54HG006434.

Author information

Authors and Affiliations

  1. CDI Laboratories, Guanajibo Research and Innovation Park, Mayaguez, PR, USA

    Pedro Ramos-López, José Irizarry & Ignacio Pino

  2. Solomon H. Snyder Department of Neuroscience, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  3. Department of Neurology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  4. Department of Ophthalmology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  5. Center for Human Systems Biology, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

  6. Institute for Cell Engineering, Johns Hopkins University School of Medicine, Baltimore, MD, USA

    Seth Blackshaw

Authors
  1. Pedro Ramos-López
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  2. José Irizarry
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  3. Ignacio Pino
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  4. Seth Blackshaw
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Corresponding author

Correspondence to Seth Blackshaw .

Editor information

Editors and Affiliations

  1. KTH School of Engineering Sciences in Chemistry, Biotechnology and Health, Protein Technology, Stockholm, Sweden

    Johan Rockberg

  2. Donnelly Centre for Cellular and Biomolecular Research, University of Toronto, Toronto, Ontario, Canada

    Johan Nilvebrant

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© 2018 Springer Science+Business Media, LLC, part of Springer Nature

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Ramos-López, P., Irizarry, J., Pino, I., Blackshaw, S. (2018). Antibody Specificity Profiling Using Protein Microarrays. In: Rockberg, J., Nilvebrant, J. (eds) Epitope Mapping Protocols. Methods in Molecular Biology, vol 1785. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7841-0_14

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  • DOI: https://doi.org/10.1007/978-1-4939-7841-0_14

  • Published:

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7839-7

  • Online ISBN: 978-1-4939-7841-0

  • eBook Packages: Springer Protocols

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