Preparation of a cyanine-based fluorescent probe for highly selective detection of glutathione and its use in living cells and tissues of mice

Jun Yin; Younghee Kwon; Dabin Kim; Dayoung Lee; Gyoungmi Kim; Ying Hu; Ji Hwan Ryu; Juyoung Yoon

doi:10.1038/nprot.2015.109

Preparation of a cyanine-based fluorescent probe for highly selective detection of glutathione and its use in living cells and tissues of mice

Jun Yin, Younghee Kwon, Dabin Kim, Dayoung Lee, Gyoungmi Kim, Ying Hu, Ji Hwan Ryu, Juyoung Yoon

Department of Chemistry & Nanoscience

Research output: Contribution to journal › Article › peer-review

96 Scopus citations

Abstract

Glutathione (GSH) is a major endogenous antioxidant that has a central role in cellular defense against toxins and free radicals. This protocol describes the preparation of CPDSA, a cyanine-based near-infrared (NIR) fluorescent probe for the detection of GSH in cells and in vivo. CPDSA is prepared with high yield through a simple two-step process. The first step is to react commercially available IR-780 iodide with excess anhydrous piperazine in anhydrous N,N-dimethyl formamide at 85 °C to form cyanine-piperazine (CP). The second step is the sulfonylation of CP with dansyl chloride in anhydrous dichloromethane. CPDSA selectively detects GSH in cells, and it has been shown to not react with other biothiols such as cysteine (Cys) and homocysteine (Hcy). This probe can also be used to monitor the GSH level of mouse bone marrow-derived neutrophils (BMDNs). The preparation of probe CPDSA takes 2 d, and experiments in cells and mice take 12-13 d.

Original language	English
Pages (from-to)	1742-1754
Number of pages	13
Journal	Nature Protocols
Volume	10
Issue number	11
DOIs	https://doi.org/10.1038/nprot.2015.109
State	Published - 1 Nov 2015

Bibliographical note

Funding Information:
acknoWleDGMents This research was supported by a grant from the National Creative Research Initiative programs of the National Research Foundation of Korea (NRF) funded by the Korean government (grant no. 2012R1A3A2048814), by the Basic Science Research Program through the NRF funded by the Ministry of Education (grant no. 2013R1A1A2008511), and by the Yonsei University College of Medicine (grant no. 6-2014-0135 to J.-H.R.) and the National Natural Science Foundation of China (grant no. 21402057).

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title = "Preparation of a cyanine-based fluorescent probe for highly selective detection of glutathione and its use in living cells and tissues of mice",

abstract = "Glutathione (GSH) is a major endogenous antioxidant that has a central role in cellular defense against toxins and free radicals. This protocol describes the preparation of CPDSA, a cyanine-based near-infrared (NIR) fluorescent probe for the detection of GSH in cells and in vivo. CPDSA is prepared with high yield through a simple two-step process. The first step is to react commercially available IR-780 iodide with excess anhydrous piperazine in anhydrous N,N-dimethyl formamide at 85 °C to form cyanine-piperazine (CP). The second step is the sulfonylation of CP with dansyl chloride in anhydrous dichloromethane. CPDSA selectively detects GSH in cells, and it has been shown to not react with other biothiols such as cysteine (Cys) and homocysteine (Hcy). This probe can also be used to monitor the GSH level of mouse bone marrow-derived neutrophils (BMDNs). The preparation of probe CPDSA takes 2 d, and experiments in cells and mice take 12-13 d.",

author = "Jun Yin and Younghee Kwon and Dabin Kim and Dayoung Lee and Gyoungmi Kim and Ying Hu and Ryu, {Ji Hwan} and Juyoung Yoon",

note = "Funding Information: acknoWleDGMents This research was supported by a grant from the National Creative Research Initiative programs of the National Research Foundation of Korea (NRF) funded by the Korean government (grant no. 2012R1A3A2048814), by the Basic Science Research Program through the NRF funded by the Ministry of Education (grant no. 2013R1A1A2008511), and by the Yonsei University College of Medicine (grant no. 6-2014-0135 to J.-H.R.) and the National Natural Science Foundation of China (grant no. 21402057).",

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AU - Hu, Ying

AU - Ryu, Ji Hwan

AU - Yoon, Juyoung

N1 - Funding Information: acknoWleDGMents This research was supported by a grant from the National Creative Research Initiative programs of the National Research Foundation of Korea (NRF) funded by the Korean government (grant no. 2012R1A3A2048814), by the Basic Science Research Program through the NRF funded by the Ministry of Education (grant no. 2013R1A1A2008511), and by the Yonsei University College of Medicine (grant no. 6-2014-0135 to J.-H.R.) and the National Natural Science Foundation of China (grant no. 21402057).

PY - 2015/11/1

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N2 - Glutathione (GSH) is a major endogenous antioxidant that has a central role in cellular defense against toxins and free radicals. This protocol describes the preparation of CPDSA, a cyanine-based near-infrared (NIR) fluorescent probe for the detection of GSH in cells and in vivo. CPDSA is prepared with high yield through a simple two-step process. The first step is to react commercially available IR-780 iodide with excess anhydrous piperazine in anhydrous N,N-dimethyl formamide at 85 °C to form cyanine-piperazine (CP). The second step is the sulfonylation of CP with dansyl chloride in anhydrous dichloromethane. CPDSA selectively detects GSH in cells, and it has been shown to not react with other biothiols such as cysteine (Cys) and homocysteine (Hcy). This probe can also be used to monitor the GSH level of mouse bone marrow-derived neutrophils (BMDNs). The preparation of probe CPDSA takes 2 d, and experiments in cells and mice take 12-13 d.

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Preparation of a cyanine-based fluorescent probe for highly selective detection of glutathione and its use in living cells and tissues of mice

Abstract

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