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Western Blotting pp 197–226Cite as

Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple Proteins

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Part of the book series: Methods in Molecular Biology ((MIMB,volume 1312))

Abstract

The qualitative and quantitative measurements of protein abundance and modification states are essential in understanding their functions in diverse cellular processes. Typical Western blotting, though sensitive, is prone to produce substantial errors and is not readily adapted to high-throughput technologies. Multistrip Western blotting is a modified immunoblotting procedure based on simultaneous electrophoretic transfer of proteins from multiple strips of polyacrylamide gels to a single membrane sheet. In comparison with the conventional technique, Multistrip Western blotting increases data output per single blotting cycle up to tenfold; allows concurrent measurement of up to nine different total and/or posttranslationally modified protein expression obtained from the same loading of the sample; and substantially improves the data accuracy by reducing immunoblotting-derived signal errors. This approach enables statistically reliable comparison of different or repeated sets of data and therefore is advantageous to apply in biomedical diagnostics, systems biology, and cell signaling research.

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Acknowledgments

The authors gratefully acknowledge Dr. Boris N Kholodenko for his support. This work was supported by National Institutes of Health Grants GM59570, AA018873, AA017261, AA007463, and AA022417.

Author information

Authors and Affiliations

  1. Department of Otolaryngology-Head and Neck Surgery, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA, 19107, USA

    Edita Aksamitiene Ph.D.

  2. Department of Pathology, Anatomy and Cell Biology, Thomas Jefferson University, 1020 Locust Street, Philadelphia, PA, 19107, USA

    Edita Aksamitiene Ph.D. & Jan B. Hoek Ph.D.

  3. Department of Biochemistry & Biophysics, Perelman School of Medicine, University of Pennsylvania, 415 Curie Blvd, Philadelphia, PA, 19104, USA

    Anatoly Kiyatkin Ph.D.

Authors
  1. Edita Aksamitiene Ph.D.
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  2. Jan B. Hoek Ph.D.
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  3. Anatoly Kiyatkin Ph.D.
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Corresponding author

Correspondence to Anatoly Kiyatkin Ph.D. .

Editor information

Editors and Affiliations

  1. Department of Medicine, University of Oklahoma, Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Health Sciences Center, Department of Veterans, Oklahoma City, Oklahoma, USA

    Biji T. Kurien

  2. Department of Medicine, Arthritis and Clinical Immunology, Oklahoma Medical Research Foundation, Health Sciences Center, Department of Veterans Affairs Medical Center, Oklahoma City, Oklahoma, USA

    R. Hal Scofield

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Aksamitiene, E., Hoek, J.B., Kiyatkin, A. (2015). Multistrip Western Blotting: A Tool for Comparative Quantitative Analysis of Multiple Proteins. In: Kurien, B., Scofield, R. (eds) Western Blotting. Methods in Molecular Biology, vol 1312. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-2694-7_23

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  • DOI: https://doi.org/10.1007/978-1-4939-2694-7_23

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-2693-0

  • Online ISBN: 978-1-4939-2694-7

  • eBook Packages: Springer Protocols

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