Immunocytochemistry/Immunofluorescence analysis using Mouse Anti-Alpha Synuclein Monoclonal Antibody, Clone 3F8 . Tissue: Primary hippocampal neurons treated with active Alpha Synuclein Protein Aggregate (SPR-322) at 4 µg/ml to induce fibrils. Species: Rat. Fixation: 4% paraformaldehyde. Primary Antibody: Mouse Anti-Alpha Synuclein Monoclonal Antibody at 1:200 for 24 hours at 4°C. Secondary Antibody: Goat Anti-Mouse Alexa Fluor 488 at 1:700 for 1 hour at RT. Counterstain: Guinea Pig Anti-NeuN (red) neuronal marker (Donkey Anti-Guinea Pig Alexa Fluor 647 1:700); DAPI (blue) nuclear stain at 1:6000, 1:3000 for 60 min at RT, 5 min at RT. Magnification: 20X. (A) DAPI (blue) nuclear stain. (B) NeuN neuronal marker (red). (C) Alpha Synuclein Antibody. (D) Composite.

Western Blot analysis of Human, Mouse, Rat Brain showing detection of 14 kDa Alpha Synuclein protein using Mouse Anti-Alpha Synuclein Monoclonal Antibody, Clone 3F8 . Lane 1: Molecular Weight Ladder (MW). Lane 2: Mouse Brain cell lysate. Lane 3: Rat brain cell lysate. Lane 4: Human brain cell lysate. Load: 15 µg. Block: 5% Skim Milk in 1X TBST. Primary Antibody: Mouse Anti-Alpha Synuclein Monoclonal Antibody at 1:1000 for 2 hours at RT. Secondary Antibody: Goat Anti-Mouse HRP:IgG at 1:3000 for 1 hour at RT. Color Development: ECL solution (Super Signal West Pico) for 5 min in RT. Predicted/Observed Size: 14 kDa.