Sensitive spectroscopic detection of large and denatured protein aggregates in solution by use of the fluorescent dye nile red

M. Sutter, S. Oliveira, N.N. Sanders, B. Lucas, A. van Hoek, M.A. Hink, A.J.W.G. Visser, S.C. De Smedt, W.E. Hennink, W. Jiskoot

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Abstract

The fluorescent dye Nile red was used as a probe for the sensitive detection of large, denatured aggregates of the model protein β-galactosidase (E. coli) in solution. Aggregates were formed by irreversible heat denaturation of β-galactosidase below and above the protein's unfolding temperature of 57.4°C, and the presence of aggregates in heated solutions was confirmed by static light scattering. Interaction of Nile red with β-galactosidase aggregates led to a shift of the emission maximum (λ
Original languageEnglish
Pages (from-to)181-192
JournalJournal of Fluorescence
Volume17
DOIs
Publication statusPublished - 2007

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