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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

Authors
 Jun Yin  ;  Younghee Kwon  ;  Dabin Kim  ;  Dayoung Lee  ;  Gyoungmi Kim  ;  Ying Hu  ;  Ji-Hwan Ryu  ;  Juyoung Yoon 
Citation
 Nature Protocols, Vol.10(11) : 1742-1754, 2015 
Journal Title
 Nature Protocols 
ISSN
 1754-2189 
Issue Date
2015
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.
URI
https://ir.ymlib.yonsei.ac.kr/handle/22282913/141543
DOI
10.1038/nprot.2015.109
Appears in Collections:
1. Journal Papers (연구논문) > 5. Research Institutes (연구소) > Research Center for Human Natural Defense System (생체방어연구센터)
Yonsei Authors
유지환(Ryu, Ji Hwan)
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Full Text
http://www.nature.com/nprot/journal/v10/n11/full/nprot.2015.109.html
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