Effect of acidic and basic pH on Thioflavin T absorbance and fluorescence

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dc.contributor.author Hackl, E. V. en
dc.contributor.author Darkwah, Joseph en
dc.contributor.author Smith, Geoff en
dc.contributor.author Ermolina, I. en
dc.date.accessioned 2016-11-10T11:45:01Z
dc.date.available 2016-11-10T11:45:01Z
dc.date.issued 2015-03-22
dc.identifier.citation Hackl, E.V., Darkwah, J., Smith, G. and Ermolina, I. (2015) Effect of acidic and basic pH on Thioflavin T absorbance and fluorescence, European Biophysics Journal, 44, (4), pp. 249-261 en
dc.identifier.uri http://hdl.handle.net/2086/12795
dc.description The file attached to this record is the author's final peer reviewed version. The Publisher's final version can be found by following the DOI link. en
dc.description.abstract Thioflavin T (ThT) is a fluorescent dye able to enhance significantly its fluorescence quantum yield upon binding to protein amyloids. ThT assay is widely used to detect and quantify amyloids in a variety of conditions, including solutions with different pH levels. In the present work, the effect of acidic and basic pH on the conformation of the ThT molecule and its absorption and fluorescence properties was studied. The results show that both acidic and basic pH decrease significantly the intensity of ThT absorption in the visible region and fluorescence emission intensity. Low pHs induce an immediate “all-or-nothing” decrease in the ThT signal, while in alkaline solutions the ThT signal decreases gradually over time. pH-induced signal quenching is less in the presence of glycerol or protein aggregates. Two different mechanisms are responsible for the ThT signal quenching—the ThT hydroxylation at basic pH and protonation of the nitrogen atom of the dimethylamino group at acidic pH. ThT assays should be carefully carried out at basic or acidic pH as strong pH dependence of ThT could be responsible for misinterpretation and false positive/negative experimental results. The potential unsuitability of ThT as a probe in solutions with high pH (>9) has been shown. © 2015, European Biophysical Societies' Association. en
dc.language.iso en en
dc.publisher Springer en
dc.subject Absorbance en
dc.subject Fluorescence en
dc.subject NMR en
dc.subject pH en
dc.subject Protein aggregation en
dc.subject ThT en
dc.title Effect of acidic and basic pH on Thioflavin T absorbance and fluorescence en
dc.type Article en
dc.identifier.doi http://dx.doi.org/10.1007/s00249-015-1019-8
dc.researchgroup Pharmaceutical Technologies en
dc.peerreviewed Yes en
dc.funder N/A en
dc.projectid N/A en
dc.cclicence CC-BY-NC en


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