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.
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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
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