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Aggregation assay

SC Simone M. Crivelli
QL Qian Luo
JS Jo A.A. Stevens
CG Caterina Giovagnoni
DK Daan van Kruining
GB Gerard Bode
SH Sandra den Hoedt
BH Barbara Hobo
AS Anna-Lena Scheithauer
JW Jochen Walter
MM Monique T. Mulder
CE Christopher Exley
MM Matthew Mold
MM Michelle M. Mielke
HV Helga E. De Vries
KW Kristiaan Wouters
DH Daniel L. A. van den Hove
DB Dusan Berkes
ML María Dolores Ledesma
JV Joost Verhaagen
ML Mario Losen
EB Erhard Bieberich
PM Pilar Martinez-Martinez
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Aβ1–42 was purchased from Anaspec. The peptide was solubilized in sterilized PBS, 0.1% trifluoroacetic acid (TFA) at the concentration 2 mM and frozen in aliquots at − 80 °C. Aliquots were diluted at the final concentration of 20 μM in a total volume of 400 μL containing 1 or 2.5 μM of affinity-purified recombinant CERTL [29]. Samples were kept under rotarod shacking for 1, 2, 4, 8, 12, and 24 h at 37 °C before adding 5 μL to 95 μL of thioflavin T (ThT) concentrated 20 μM, dispensed in 96-well optical plate and measuring fluorescent excitation at 450 nm and emission 486 nm in Victor X3 plate reader (Perkin-Elmer). Two Aβ antibodies against epitope 1–16 (6E10, Biolegend) and 17–24 (4G8, Biolegend) respectively were used to antagonize aggregation at a concentration of 0.1 mg/ml.

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