Anti-alpha Synuclein (phospho Ser129) Antibody [J18] (A304933)

Immunocytochemistry/Immunofluorescence analysis of human iPSC-derived neurons, using Anti-alpha Synuclein (phospho Ser129) Antibody [J18] (A304933), at 1:1,000 for O/N at 4°C. The secondary antibody used was Anti-Rabbit: A488 at 1:1,000 for 1 hour at room temperature. Magnification: 40X. Nuclear stain: Hoechst- 20 min, room temperature (blue). Actin stain: Phalloidin-647- 20 min, room temperature (magenta). 4K cells per well. iPSC neurons: Applied StemCell Catalog # ASE-9321K. A) negative control; no fibrils added to well. B) 7 days after addition of active recombinant human pre-formed fibrils (Type 1). StressMarq catalog # SPR-322. Sonicated before use, 2.5 ug per well.

Western blot analysis of mouse brain showing detection of Alpha Synuclein pSer129 protein using Anti-alpha Synuclein (phospho Ser129) Antibody [J18] (A304933) at 1:500 for 2 hours at room temperature with shaking. Lane 1: MW ladder. Lane 2: mouse brain. Load: 15 ug. Block: 5% BSA in TBST. The secondary antibody used was Goat anti-rabbit IgG:HRP at 1:4,000 for 1 hour at room temperature with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 minutes in room temperature.

Dot blot analysis of alpha synuclein peptide using Anti-alpha Synuclein (phospho Ser129) Antibody [J18] (A304933) at 1:500 for 2 hours at room temperature with shaking. The secondary antibody used was Goat anti-rabbit IgG:HRP at 1:4,000 for 1 hour at room temperature with shaking. Phospho peptide sequence: AYEMP-pS-EEGYQ. Non-phospho peptide sequence: AYEMPSEEGYQ. This sequence is the same for human, mouse, and rat.

Western blot analysis of human Alpha Synuclein showing detection of Alpha Synuclein pSer129 protein using Anti-alpha Synuclein (phospho Ser129) Antibody [J18] (A304933) at 1:500 for 2 hours at room temperature with shaking. Lane 6: 15 ug human Parkinson€™s Disease brain lysate.. Block: 5% BSA in TBST. The secondary antibody used was Goat anti-rabbit IgG:HRP at 1:4,000 for 1 hour at room temperature with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 minutes in room temperature.