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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>"}],"abstractLicenseFlag":"disallow"}],"creator":[{"@id":"https://cir.nii.ac.jp/crid/1410001205223548293","@type":"Researcher","personIdentifier":[{"@type":"NRID","@value":"9000240058836"}],"foaf:name":[{"@language":"en","@value":"Tomisawa Satoshi"}],"jpcoar:affiliationName":[{"@language":"en","@value":"Faculty of Advanced Life Science, Hokkaido University"}]},{"@id":"https://cir.nii.ac.jp/crid/1410001205223548291","@type":"Researcher","personIdentifier":[{"@type":"NRID","@value":"9000240058837"}],"foaf:name":[{"@language":"en","@value":"Abe Chiharu"}],"jpcoar:affiliationName":[{"@language":"en","@value":"Faculty of Advanced Life Science, Hokkaido 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