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Sensitive detection of fluorescence in western blotting by merging images

by Yukari Kondo, Shinichiro Higa, Takeshi Iwasaki, Tomoya Matsumoto, Kazumitsu Maehara, Akihito Harada, Yoshihiro Baba, Masatoshi Fujita, Yasuyuki Ohkawa

The western blotting technique is widely used to analyze protein expression levels and protein molecular weight. The chemiluminescence method is mainly used for detection due to its high sensitivity and ease of manipulation, but it is unsuitable for detailed analyses because it cannot be used to detect multiple proteins simultaneously. Recently, more attention has been paid to the fluorescence detection method because it is more quantitative and is suitable for the detection of multiple proteins simultaneously. However, fluorescence detection can be limited by poor image resolution and low detection sensitivity. Here, we describe a method to detect fluorescence in western blots using fluorescence microscopy to obtain high-resolution images. In this method, filters and fluorescent dyes are optimized to enhance detection sensitivity to a level similar to that of the chemiluminescence method.

Authors:   Yukari Kondo; Shinichiro Higa; Takeshi Iwasaki; Tomoya Matsumoto; Kazumitsu Maehara; Akihito Harada; Yoshihiro Baba; Masatoshi Fujita; Yasuyuki Ohkawa
Journal:   PLoS ONE
Volume:   13
edition:   1
Year:   2018
Pages:   e0191532
DOI:   10.1371/journal.pone.0191532
Publication date:   19-Jan-2018
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