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

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

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 s...

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Veröffentlicht in:BUNSEKI KAGAKU 2019/12/05, Vol.68(12), pp.961-964
Hauptverfasser: MORI, Takeshi, KAMINO, Kenta, ODA, Tsuyoshi, KATAYAMA, Yoshiki
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Sprache:eng ; jpn
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flow cytometry
fluorescence
membrane protein
single cell
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container_issue 12
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container_title BUNSEKI KAGAKU
container_volume 68
creator MORI, Takeshi
KAMINO, Kenta
ODA, Tsuyoshi
KATAYAMA, Yoshiki
description 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.
doi_str_mv 10.2116/bunsekikagaku.68.961
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subjects flow cytometry
fluorescence
membrane protein
single cell
title Improvement of the Signal-to-noise Ratio on the Fluorescent Detection of a Membrane Protein Using Horse Radish Peroxidase
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