A new approach to detect small peptides clearly and sensitively by Western blotting using a vacuum-assisted detection method

DOI Web Site PubMed 被引用文献1件 参考文献9件 オープンアクセス

書誌事項

公開日
2013
資源種別
journal article
DOI
  • 10.2142/biophysics.9.79
公開者
日本生物物理学会

説明

Western blotting is a widely used technique for the detection and quantification of proteins and peptides. However, it is challenging to detect small peptides efficiently by the conventional Western blotting method with shaking, in part because the peptides readily detach from the blotted membrane. Although some modified Western blotting protocols have been developed to overcome this problem, it remains difficult to prevent peptide detachment from the membrane. In this study, we show that the previously developed vacuum-assisted detection method greatly improves the detection of small peptides without additional protocol modification. The vacuum-assisted method was developed to shorten the time required for all immunodetection steps, and all the Western blotting solutions penetrated the membrane quickly and efficiently by this method. By using this vacuum method, we succeeded in detecting small peptides that were completely undetectable by the conventional Western blotting method. We also confirmed that peptide detachment was induced even by gentle shaking in the case of the conventional method, and the detachment was accelerated when detergent was present in the buffer. Unlike in the conventional method, there is no need to shake the membrane in solution in the vacuum method. Therefore, it is thought that the small peptides could be detected sensitively only by the vacuum method.<br>

収録刊行物

  • BIOPHYSICS

    BIOPHYSICS 9 (0), 79-83, 2013

    日本生物物理学会

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