Fibril formation monitored by ThT.<b> </b><b>aggregation</b><b> </b><b>of amyloid</b><b> </b><b>b</b><b> </b><b>by increasing A</b><b>β</b><b> </b><b>dynamics</b>

The Aβ monomer was prepared from 26-<i>O</i>-acyliso-Aβ₄₂ (iso-Aβ), which starts aggregating immediately after dissolving in an aqueous solution. Formation of Aβ aggregates in the presence of 10 mM ATP, ADP or AMP and the absence were monitored by ThT fluorescence. The aggregation proceeded in a final concentration of 10 % v/v DMSO. The fluorescence of 10 μM ThT in the presence or absence of 2.2 μM Aβ₄₂ was monitored at an excitation wavelength of 444 nm and emission wavelength of 485 nm.本研究所用β淀粉样蛋白单体（Aβ monomer）由26-O-酰基异β淀粉样蛋白42肽（26-O-acyliso-Aβ₄₂，简称iso-Aβ）制备得到，该前体在水溶液中溶解后会立即启动聚集过程。在10 mM三磷酸腺苷（ATP）、二磷酸腺苷（ADP）或一磷酸腺苷（AMP）存在，以及不存在这些核苷酸的条件下，通过硫黄素T（Thioflavin T，ThT）荧光法监测β淀粉样蛋白聚集体的形成过程。聚集反应在终体系中含有10%（v/v）二甲基亚砜（DMSO）的条件下进行。针对含与不含2.2 μM β淀粉样蛋白42肽（Aβ₄₂）的体系，对10 μM硫黄素T的荧光信号进行监测，检测时激发波长设为444 nm，发射波长设为485 nm。