Kinetics of Amyloid-β Aggregation

The thioflavine T (ThT) assay was used to measure β-sheet formation over time with a fluorescence read-out. Different concentrations (0, 10, 20, 100 μM) of Aβ_1-42 and scrambled Aβ_1-42 were added to non-binding 96-well microplates (655,906, Greiner) with 12 μM of ThT in PBS with 1 mM EDTA. Fluorescence was measured at 5 min-intervals for 8 h on a plate reader (Victor 31,420 Multilabel Counter, Perkin Elmer) at 21°C using excitation and emission wavelengths of 440 nm and 480 nm, respectively, with an automated protocol.

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Vande Vyver M., Daeninck L., De Smet G., Aourz N., Sahu S., Engelborghs S., Pauwels K., De Bundel D, & Smolders I. (2023). The intracerebral injection of Aβ1-42 oligomers does not invariably alter seizure susceptibility in mice. Frontiers in Aging Neuroscience, 15, 1239140.

Publication 2023

Victor 31,420 Multilabel Counter

Assay Edta Fluorescence Thioflavine t

Corresponding Organization : Vrije Universiteit Brussel

Other organizations : University of Antwerp, Universitair Ziekenhuis Brussel

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Variable analysis

independent variables

Aβ1-42 concentrations (0, 10, 20, 100 μM)
Scrambled Aβ1-42 concentrations (0, 10, 20, 100 μM)

dependent variables

β-sheet formation over time measured by ThT fluorescence

control variables

ThT concentration (12 μM)
Buffer (PBS with 1 mM EDTA)
Temperature (21°C)
Excitation and emission wavelengths (440 nm and 480 nm, respectively)
Microplate used (non-binding 96-well microplates, 655,906, Greiner)

controls

Positive control: Aβ1-42 samples
Negative control: Scrambled Aβ1-42 samples

Annotations

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