Improvement of the Signal-to-noise Ratio on the Fluorescent Detection of a Membrane Protein Using Horse Radish Peroxidase

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  • MORI Takeshi
    Department of Applied Chemistry, Faculty of Engineering, Kyushu University Graduate School of Systems Life Sciences, Kyushu University
  • KAMINO Kenta
    Graduate School of Systems Life Sciences, Kyushu University
  • ODA Tsuyoshi
    Graduate School of Systems Life Sciences, Kyushu University
  • KATAYAMA Yoshiki
    Department of Applied Chemistry, Faculty of Engineering, Kyushu University Graduate School of Systems Life Sciences, Kyushu University Center for Advanced Medical Innovation, Kyushu University Department of Biomedical Engineering, Chung Yuan Christian University

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Other Title
  • 西洋ワサビペルオキシダーゼを用いる膜タンパク質の蛍光標識における信号対雑音比の改善
  • セイヨウ ワサビペルオキシダーゼ オ モチイル マク タンパクシツ ノ ケイコウ ヒョウシキ ニ オケル シンゴウ タイ ザツオンヒ ノ カイゼン

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Abstract

<p>Horseradish peroxidase (HRP) is used as a sensitizing enzyme to detect a membrane protein with high sensitivity. In this method, the membrane protein is labeled with HRP, and fluorescent group-modified tyramide is used as a substrate for fluorescent signal amplification. However, a high background signal due to a nonspecific adsorption of fluorescent group-modified tyramide to cells is known to compromise high-sensitive detection. Here, we designed a new substrate in which tyramide was changed to tyrosyltaurine. Because the background signal was greatly reduced by the sulfonate group in the new substrate, the signal-to-noise ratio was greatly improved compared with a commercial substrate.</p>

Journal

  • BUNSEKI KAGAKU

    BUNSEKI KAGAKU 68 (12), 961-964, 2019-12-05

    The Japan Society for Analytical Chemistry

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