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The thioflavin strongTstrong fluorescence assay for amyloid fibril.pdf

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University of Wollongong Research Online Faculty of Science - Papers (Archive) Faculty of Science, Medicine and Health 2009 The thioflavin T fluorescence assay for amyloid fibril detection can be biased by the presence of exogenous compounds Sean A . Hudson University of A delaide Heath Ecroyd University of Wollongo ng, heathe@.au Tak W. Kee University of A delaide J ohn A. Carver Publication Details Hudson, S. A., Ecroyd, H., Kee, T. W. Carver, J. A. (2009). The thioflavin T fluorescence assay for amyloid fibril detection can be biased by the presence of exogenous compounds. Febs Journal, 276 (20), 5960-5972. esearch Online is the open access institutional repository for the University of Wollongong. For further information contact the UOW Library: research-pubs@.au The thioflavin T fluorescence assay for amyloid fibril detection can be biased by the presence of exogenous compounds Abstract Thioflavin T (ThT) dye fluorescence is used regularly to quantify the formation and inhibition of amyloid fibrils in the presence of anti-amyloidogenic compounds such as polyphenols. However, in this study, it was shown, using three polyphenolics (curcumin, quercetin and resveratrol), that ThT fluorescence should be used with caution in the presence of such exogenous compounds. The strong absorptive and fluorescent properties of quercetin and curcumin were found to significantly bias the ThT fluorescence readings in both in situ real- time ThT assays and single time-point dilution ThT-type assays. The presence of curcumin at concentrations as low as 0.01 and 1 uM was sufficient to interfere with the ThT fluorescence associated with fibrillar amyloid- b(1-42) (0.5 uM) and fibrillar reduced and carboxymethylated kappa-casein (50 lm), respectively. The T