Ab272736

Paraffin-embedded brain tissues were sliced into 6 μm sections, and then subjected to immunofluorescence staining. After 5 min of antigen retrieval, the sections were incubated in 5% goat serum for 1 h and incubated with rabbit TH (dilution ratio of 1:500, GB11181, Servicebio, Wuhan, China) and 4-HNE (dilution ratio of 1:50, MA5-27570, Invitrogen), and α-SYN (dilution ratio of 1:500, ab272736, Abcam) at 4 °C overnight and with Cy3-conjugated donkey anti-rabbit antibody (dilution ratio of 1:250, GB 21403, Servicebio) or Goat Anti-Mouse Alexa Fluor 488 (dilution ratio of 1:1000, A32723, Invitrogen) for 1 h. A confocal laser microscope (Olympus) was employed to observe the immune-response cells.

For immunostaining, brains were sliced into 50 μm coronal slices by a vibration microtome (Leica, VT1000 S). The brain slices were washed in PBS three times at room temperature, blocked with 5% (wt/vol) BSA (bovine serum albumin) containing 0.3% Triton-X-100 (vol/vol) in 0.01 M PBS for 1.5 h at room temperature and then incubated with primary antibodies for 24 h at 4 °C. The following primary antibodies were used: anti-PV (1:500, mouse, Millipore, MAB1572), anti-synaptophysin (1:200, mouse, Abcam, ab8049), anti-synapsin (1:500, rabbit, Abcam, ab254349), anti-SV2 (1:500, rabbit, Abcam, ab32942), anti-SNAP-25 (1:100, rabbit, Abcam, ab108990), anti-syntaxin (1:100, rabbit, Abcam, ab272736), anti-beta amyloid (1:800, mouse, Abcam, ab126649), and anti-GFP (1:500 rabbit, Abcam, ab290). Then, the sections were washed in PBS four times for 15 min and incubated with the corresponding secondary antibodies for 1.5 h at room temperature. All antibodies were diluted in the same block solution.