Fig. 5: Different α-synuclein species are present in different sucrose fractions.

A Principle of the single-molecule pulldown (SiMPull) assay using the same pair of antibodies (abbreviated as ab in the schematic), including representative TIRF images for the 50% fraction using the MJF and SC antibodies. The biotinylated capture antibody (abbreviated as ab in the schematic) is attached to the passivated surface via a biotin (orange) neutravidin linkage (grey). The capture antibody can pulldown its distinct antigen. The captured aggregates are sandwiched by primary monoclonal fluorescently labelled (red star) detection antibody. Scale bar = 5 µm. The images are contrast adjusted. The schematic has been created with BioRender.com. B Number of detected species in each sucrose fraction using the conformation-specific MJF antibody targeting aggregated filamentous α-synuclein. Error bars are mean ± STD from 15 different fields of view. Each individual point is representing one field of view. C Number of detected aggregates in each species using the sequence-specific SC epitope-specific antibody targeting amino acids 121–125. Error bars are mean ± STD from 15 different fields of view. Each individual point is representing one field of view. One representative replicate out of three is displayed here. Each individual colour is representing a distinct sucrose fraction (grey = 10%, orange = 20%, green = 30%, blue = 40%, violet = 50%). Source data are provided as a Source Data file. MJF = MJFR-14-6-4-2 antibody, SC = Santa cCruz 211 antibody, μm = Micrometer, # = Number. Details about the antibodies can be found in Supplementary Table 3.