Thioflavin T Assay for Amyloid Detection

ApoAI-M, apoAI-WT and apoAI-Iowa(G26R) variant (0.2 mg/ml) were incubated at 37°C and diluted with ThT stock solution at time of use. 180 µl of protein was incubated for 10 min in the dark with 20 µl of a ThT (100 µM)/glycine (10 mM) solution (ThT stock: 1 mM stored in the dark at 4°C; Glycine buffer stock: 0.1 M at pH 8.5 stored at 4°C). ThT fluorescence was then measured using a VICTOR3 Multilabel Plate Counter (PerkinElmer, Waltham, MA, USA) spectrofluorometer at an excitation wavelength of 450 nm and an emission wavelength of 545 nm, with excitation and emission slit widths of 10 nm [16] (link).