Immunoprecipitation of Amyloid Fibrils by the Use of an Antibody that Recognizes a Generic Epitope Common to Amyloid Fibrils
Figure 5
Immunoprecipitation of amyloid fibrils using the LOC antibody.
(A) Schematic of the protocol used to isolate amyloid fibrils. (B) Aβ1–40, (C) gelsolin or (D) α-syn amyloid fibrils (0.2% w/w) were added to worm PDS, digested with PK for 2 h at 42°C and precipitated with 1 volume of cold acetone. The pellet was resuspended in buffer containing LOC antibody and the IP was performed as described in Materials and Methods. As a negative control, we performed the IP in the absence of LOC antibody (beads). The samples were resolved by SDS-PAGE (16% tris-tricine gels) and probed for Aβ1–40, gelsolin, or α-syn by western blotting.