Fluorescent Antibody-based Detection of Protein Aggregates

Fluorescent antibodies were used for microscopy-based detection of aggregates. The mouse monoclonal anti-α-synuclein Syn211 antibody (Santa Cruz Biotechnology, Inc., Dallas, USA) was labeled with CF633 and CF488A (Biotium, Freemont, USA) and the anti-Aβ IC16 (Heinrich-Heine University, Düsseldorf, Germany) antibody was labeled with CF633 (Biotium, Freemont, USA)^{40 (link),44 (link)}. The labeling process was performed according to the manufacturer’s protocol. The dyes with activated succinimidyl esters were mixed with the antibody in carbonate buffer to react covalently with the amines of the antibody. The labeled antibody was purified using a polyacrylamide bead suspension (Bio-Gel P 30 Gel, Bio-Rad Laboratories, Inc., Hercules, USA). The concentration and degree of labeling were determined according to the manufacturer’s protocol.