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On-Membrane Deblocking of Proteins

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Part of the book series: Methods in Molecular Biology™ ((MIMB,volume 64))

Abstract

Vandekerckhove et al. (1) developed a new method for protein micro-sequencing in 1985, in which subnanomole amounts of proteins are separated by one-dimensional (1D) or two-dimensional (2D) polyacrylamide gel electrophoresis (PAGE) and then transferred from the gel onto a Polybrene-treated glass fiber filter by Western blotting originally developed by Towbin et al. (2) and sequenced directly by a gas-phase sequencer (3). Two years later, Matsudaira (4) found that a polyvinylidene difluoride (PVDF) membrane could be used in place of the glass fiber filter for efficient electroblotting and gas-phase sequencing. The PVDF membrane filter has several advantages over the glass fiber filter in, for example, protein binding capacity, protein detection on the filter, and handling. Since 1987, the Western blotting/sequencing using the PVDF membrane has come to be very widely used in protein sequence analysis (5).

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References

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© 1997 Humana Press Inc.

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Hirano, H., Komatsu, S., Tsunasawa, S. (1997). On-Membrane Deblocking of Proteins. In: Smith, B.J. (eds) Protein Sequencing Protocols. Methods in Molecular Biology™, vol 64. Humana Press, Totowa, NJ. https://doi.org/10.1385/0-89603-353-8:285

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  • DOI: https://doi.org/10.1385/0-89603-353-8:285

  • Publisher Name: Humana Press, Totowa, NJ

  • Print ISBN: 978-0-89603-353-5

  • Online ISBN: 978-1-59259-550-1

  • eBook Packages: Springer Protocols

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