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An Overview of Western Blotting for Determining Antibody Specificities for Immunohistochemistry

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Signal Transduction Immunohistochemistry

Part of the book series: Methods in Molecular Biology ((MIMB,volume 717))

Abstract

Despite its overall simplicity, protein blotting or Western blotting has been proven to be a powerful procedure for the immunodetection of proteins, especially those that are of low abundance, following electrophoresis. 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 since its inception and researchers have a variety of ways and means to carry out this transfer. This procedure is used in combination with other important antibody-based detection methods such as enzyme-linked immunosorbant assay and immunohistochemistry to provide confirmation of results both in research and diagnostic testing. Specificity of antibodies used for immunohistochemistry is of critical importance and therefore Western blot is a “must” to address antibodies’ specificity.

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Author information

Authors and Affiliations

  1. Arthritis and Immunology Program, Oklahoma Medical Research Foundation, Oklahoma City, OK, USA

    Biji T. Kurien, Yaser Dorri, Skyler Dillon, Anil Dsouza & R. Hal Scofield

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

    Biji T. Kurien, Yaser Dorri, Skyler Dillon & Anil Dsouza

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

    R. Hal Scofield

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  1. Biji T. Kurien
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  2. Yaser Dorri
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  3. Skyler Dillon
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  4. Anil Dsouza
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  5. R. Hal Scofield
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Correspondence to Biji T. Kurien .

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  1. R & D Systems, Inc., McKinley Place NE. 614, Minneapolis, 55413, Minnesota, USA

    Alexander E. Kalyuzhny

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Kurien, B.T., Dorri, Y., Dillon, S., Dsouza, A., Scofield, R.H. (2011). An Overview of Western Blotting for Determining Antibody Specificities for Immunohistochemistry. In: Kalyuzhny, A. (eds) Signal Transduction Immunohistochemistry. Methods in Molecular Biology, vol 717. Humana Press. https://doi.org/10.1007/978-1-61779-024-9_3

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  • DOI: https://doi.org/10.1007/978-1-61779-024-9_3

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  • Publisher Name: Humana Press

  • Print ISBN: 978-1-61779-023-2

  • Online ISBN: 978-1-61779-024-9

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