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Assay Method Information

Assay Name:  Thioflavin-T (Th-T) Fluorescence Assay
Description:  Fluorescence intensity was measured at 420 nm excitation and 485 nm emission using a microplate reader (MPR-A4╬╣II; TOSOH, Tokyo, Japan, or Fluoroskan Ascent; Thermo Scientific, Rockford, IL). In brief, Aβ42 was dissolved in 0.1% NH4OH at 250 uM, and each flavonoid was dissolved in EtOH at 5 mM, followed by dilution with sodiumphosphate-buffered saline (PBS: 50 mM sodium phosphate and 100 mM NaCl, pH 7.4) at the desired concentration (Aβ42, 25 uM; flavonoids, 50 uM). NaIO4 or Tris(2-carboxyethyl)phosphine hydrochloride (TCEP-HCl) was initially dissolved in PBSat 100 mM, then diluted with PBS at 100 uM before use. Experiments under an anaerobic condition were performed in a desiccator evacuated by a diaphragm pump (about 8mmHg; KNF Lab LABOPORT vacuum pump, KNF Neuberger, NJ) at room temperature. Unless otherwise noted, the concentrations of Aβ42, flavonoids, and oxidant/reductant used in this study were 25, 50, and 100 uM, respectively.
Affinity data for this assay
 

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Last update November 1, 2007
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