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Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods

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Signal Transduction Immunohistochemistry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1554))

Abstract

Immunoblotting has been used in conjunction with other important antibody based detection methods like enzyme linked immunosorbent assay and immunohistochemistry to provide confirmation of results both in research and diagnostic testing. Specificity of antibodies employed for immunohistochemical studies is of critical importance and therefore the use of western blotting is imperative to address specificity of antibodies. In spite of its overall simplicity, western blotting or protein blotting is a powerful procedure for immunodetection of proteins, especially those that are of low abundance, following electrophoretic separation. The usefulness of this procedure stems from its ability to provide simultaneous resolution of multiple immunogenic antigens within a sample for detection by specific antibodies. Protein blotting has evolved greatly over the last few decades and researchers have a variety of ways and means to carry out this procedure to validate antibodies for immunohistochemistry.

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Author information

Authors and Affiliations

  1. Arthritis and Clinical Immunology Program, Oklahoma Medical Research Foundation, 825 NE 13th Street, Oklahoma City, OK, 73104, USA

    Biji T. Kurien & R. Hal Scofield

  2. Department of Medicine, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    Biji T. Kurien & R. Hal Scofield

  3. Department of Veterans Affairs Medical Center, University of Oklahoma Health Sciences Center, Oklahoma City, OK, 73104, USA

    Biji T. Kurien & R. Hal Scofield

Authors
  1. Biji T. Kurien
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  2. R. Hal Scofield
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Corresponding author

Correspondence to Biji T. Kurien .

Editor information

Editors and Affiliations

  1. Immunocytochemistry & Elispot Assays, Bio-Techne, Inc., Minneapolis, Minnesota, USA

    Alexander E. Kalyuzhny

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Kurien, B.T., Scofield, R.H. (2017). Validating Antibody Specificities for Immunohistochemistry by Protein Blotting Methods. In: Kalyuzhny, A. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 1554. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6759-9_3

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  • DOI: https://doi.org/10.1007/978-1-4939-6759-9_3

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-6757-5

  • Online ISBN: 978-1-4939-6759-9

  • eBook Packages: Springer Protocols

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