Journal
JOURNAL OF PHYSICAL CHEMISTRY B
Volume 112, Issue 47, Pages 15174-15181Publisher
AMER CHEMICAL SOC
DOI: 10.1021/jp805560c
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Funding
- Drug Research Academy
- Carlsberg Foundation
- Italian Ministry of University and Research [PRIN 2005]
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The fluorescent dye thioflavin T (ThT) is commonly used for in situ amyloid fibril detection. In this work, we focused on the spectroscopic properties and chemical stability of ThT in aqueous solution as a function of pH, temperature, and dye concentration. A reversible hydroxylation process occurs in alkaline solutions, which was characterized using a combination of UV-vis absorption spectroscopy, proton NMR, and density functional theory (DFT). On the basis of these studies, we propose a chemical structure for the hydroxylated form. Finally, by means of fluorescence spectroscopy, ThT hydroxylation effects on in situ amyloid detection have been investigated, providing new insights on the efficiency of the ThT assay for quantitative fibril evaluation at basic pH.
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