Ab218818

Recombinant human α-syn aggregates were purchased from Abcam (ab218819) along with the corresponding monomers (ab218818). Dopamine (HB1835) and ZD 7288 (HB1152) were purchased from Hello Bio (HB1835). Glibenclamide (PHR1287) was purchased from Sigma-Aldrich. The sheep polyclonal anti-tyrosine hydroxylase was purchased from Merck (AB1542) and donkey anti-sheep 488 secondary from Invitrogen (A11015). The Alexa Fluor 594 hydrazide dye was purchased from Invitrogen (10072752).

Differentiated SH-SY5Y cells were treated with human α-syn protein aggregates (active) (ab218819, 4 μg/ml; Abcam) and human α-syn protein monomer (active) (ab218818; Abcam). Immunostainings were performed as described previously. Differentiated SH-SY5Y cells were fixed with 4% PFA and subsequently incubated with primary antibodies against phosphorylated S129 α-syn (α-syn-P; 1:1,000; ab184674) at 4°C for 24 h. After washing 3× with PBS, the cells were incubated with Alexa Fluor 647 anti-mouse secondary antibody (1/500) for 1 h at RT, counterstained with Hoechst (blue) nuclear stain (1/4,000), and mounted on slides with antifade reagent. Images were acquired by a Nikon AIR confocal microscope using a 60× objective.