On-Membrane Deblocking of Proteins

  • Hisashi Hirano
  • Setsuko Komatsu
  • Susumu Tsunasawa
Part of the Methods in Molecular Biology™ book series (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).

Keywords

Vortex Acetonitrile Lipase Trypsin Acetyl 

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Copyright information

© Humana Press Inc. 1997

Authors and Affiliations

  • Hisashi Hirano
    • 1
  • Setsuko Komatsu
    • 1
  • Susumu Tsunasawa
    • 1
  1. 1.Kihara Institute for Biological ResearchYokohama City UniversityYokohamaJapan

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