Kondo Yukari, Higa Shinichiro, Iwasaki Takeshi, Matsumoto Tomoya, Maehara Kazumitsu, Harada Akihito, Baba Yoshihiro, Fujita Masatoshi, Ohkawa Yasuyuki
Division of Transcriptomics, Medical Institute of Bioregulation, Kyushu University, Fukuoka, Japan.
Department of Cellular Biochemistry, Graduate School of Pharmaceutical Sciences, Kyushu University, Fukuoka, Japan.
PLoS One. 2018 Jan 19;13(1):e0191532. doi: 10.1371/journal.pone.0191532. eCollection 2018.
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.
蛋白质印迹技术被广泛用于分析蛋白质表达水平和蛋白质分子量。化学发光法因其高灵敏度和易于操作而主要用于检测，但由于它不能同时检测多种蛋白质，因此不适合进行详细分析。最近，荧光检测法受到了更多关注，因为它更具定量性，并且适合同时检测多种蛋白质。然而，荧光检测可能会受到图像分辨率差和检测灵敏度低的限制。在这里，我们描述了一种使用荧光显微镜检测蛋白质印迹中荧光以获得高分辨率图像的方法。在这种方法中，对滤光片和荧光染料进行了优化，以将检测灵敏度提高到与化学发光法相似的水平。
