Thioflavin T (Basic Yellow 1)

A fluorescent probe for amyloid fibrils

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货号: ajce44304
CAS: 2390-54-7
别名: 硫黄素T; Basic Yellow 1
分子式: C17H19ClN2S
分子量: 318.86
纯度: >98%
溶解度: DMSO : 22.5 mg/mL (70.56 mM);Water : 12.5 mg/mL (39.20 mM)
储存: Store at RT, protect from light
库存: 现货

Background

Thioflavin T is a fluorescent probe for amyloid fibrils.¹ It displays excitation/emission maxima of 385/445 nm, respectively, and exhibits an increase in fluorescence intensity and a shift in excitation/emission maxima to 450/482 nm upon binding to amyloid fibrils.^1,2 The fluorescence intensity is also pH dependent, increasing in intensity as the pH increases.² Thioflavin T binds to different types of amyloid fibrils with varying affinity, over two orders of magnitude, depending on type and binding mode.³ It has been used with fluorescence microscopy to visualize brain amyloid fibril plaques in a rat model of Alzheimer's disease.⁴

1.Biancalana, M., and Koide, S.Molecular mechanism of Thioflavin-T binding to amyloid fibrilsBiochim. Biophys. Acta1804(7)1405-1412(2010) 2.Naiki, H., Higuchi, K., Hosokawa, M., et al.Fluorometric determination of amyloid fibrils in vitro using the fluorescent dye, thioflavin TAnal. Biochem.177(2)244-249(1989) 3.Sulatskaya, A.I., Kuznetsova, I.M., Belousov, M.V., et al.Stoichiometry and affinity of thioflavin T binding to Sup35p amyloid fibrilsPLoS One11(5)e0156314(2016) 4.Alimohammadi-Kamalabadi, M., Eshraghian, M., Zarindast, M.-R., et al.Effect of creatine supplementation on cognitive performance and apoptosis in a rat model of amyloid-beta-induced Alzheimer's diseaseIran J. Basic Med. Sci.19(11)1159-1165(2016)

Protocol

Kinase experiment:

Thioflavin T (ThT) is prepared by dissolving ~3 mg dry powder in 1 mL water. The solution is filtered through 0.22 μm syringe filters followed by measurement of the concentration by diluting the stock solution in ethanol and using an extinction coefficient of 26,620 M-1 cm-1 at 416 nm. The stock solution is stored at 4 °C covered with foil and used for up to a month to make assay solutions by diluting either in water or desired buffer[1]. Thioflavin T (ThT) fluorescence emission is measured with excitation at 450 nm and recording the spectrum between 465 and 565 nm with 5 nm slits using a FluoroMax 2 spectrofluorometer. The excitation spectra are collected by setting the emission wavelength to 482 nm and collecting the spectrum between 300 and 470 nm with 5 nm slit widths, and 1 s integration time and 1 nm interval. Emission spectra between 465 and 565 nm are collected upon excitation at 450 nm. Excitation and emission spectra in the presence of amyloid fibrils are measured with varying concentrations of Thioflavin T and 5 ng/mL amyloid (calculated based on starting protein concentration) before collecting the spectra[1].

参考文献:

[1]. Khurana R, et al. Mechanism of thioflavin T binding to amyloid fibrils. J Struct Biol. 2005 Sep;151(3):229-38.