Kazuhisa Hirabayashi, Dr. Kenjiro Hanaoka, Dr. Manabu Shimonishi, Takuya Terai, Dr. Toru Komatsu, Dr. Tasuku Ueno and Prof. Dr. Tetsuo Nagano Inside Cover: Selective Two-Step Labeling of Proteins with an Off/On Fluorescent Probe (Chem. Eur. J. 52/2011) Chemistry - A European Journal 17
Fluorescent Probes In their Full Paper on page 14763 ff., T. Nagano et al. describe the selective two-step labeling of proteins with an off/on fluorescent probe. The probe consists of dichlorofluorescein conjugated with nitrilotriacetic acid (NTA)-Ni2+ as the His-tag recognition site and a 2,4-dinitrophenyl ether moiety, which quenches the probe's fluorescence by photoinduced electron-transfer (PeT) from the excited fluorophore to the 2,4-dinitrophenyl ether and also has reactivity with cysteine. Removal of the 2,4 dinitrophenyl ether quencher by a proximity-enhanced reaction with the cysteine residue of the modified tag results in a marked fluorescence increase.
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