Preparation and Application of Amyloid-beta Oligomers

Soluble Aβ42 oligomers or soluble scrambled Aβ42 oligomers were prepared as described (32 (link), 104 (link)). One milligram of lyophilized human Aβ42 (Anaspec) or scrambled Aβ42 (Anaspec) was dissolved in 1 ml of 1,1,1,3,3,3-hexafluoro-2-propanol (Fisher Scientific) to prevent aggregation, portioned into 10-μg aliquots, air dried, and stored at −80 °C. For use in experiments, an aliquot was thawed at room temperature and then dissolved in 100% dimethyl sulfoxide, then diluted into PBS to make a 100 μM solution. The solution was incubated for 16 h at 4 °C and then diluted to a final concentration for use in experiments. The following agonists were used in this study: 1 μM PNU-120596 (Alomone Labs), 2 μM RJR-2403 Oxalate (Alomone labs), 1 μM NS-3861 (Tocris Bioscience), and 1 μM Carbamoylcholine chloride (carbachol) (Tocris Bioscience). Aβcore (YEVHHQ) and inactive Aβcore (SEVAAQ) peptides were prepared as described (43 (link)).