Alpha Synuclein Antibody (pSer129)

• Catalog Number: orb612713
• Category: Antibodies
• Description: Rabbit monoclonal antibody to Alpha Synuclein (Phospho-Ser129)
• Species/Host: Rabbit
• Clonality: Monoclonal
• Clone Number: J18
• Tested applications: ELISA, ICC, IHC, WB
• Reactivity: Human, Mouse
• Isotype: IgG
• Immunogen: Human alpha synuclein AA 124-134: AYEMP-pS-EEGYQ-Cys
• Concentration: 1 mg/ml
• Dilution range: WB (1:500)
• Conjugation: Unconjugated
• Target: Alpha Synuclein pSer129
• Entrez: 6622
• UniProt ID: P37840
• NCBI: NP_000336.1
• Storage: -20°C
• Buffer/Preservatives: PBS pH 7.4, 50% glycerol, 0.09% sodium azide *Storage buffer changes when conjugated
• Alternative names: Phosphorylated alpha synuclein antibody, Phospho a
• Note: For research use only
• Application notes: A 1:500 dilution of SMC-600 was sufficient for detection of Alpha Synuclein pSer129 in 10 µg of Mouse Brain by ECL immunoblot analysis using Goat Anti-Rabbit IgG:HRP as the secondary antibody.
• Expiration Date: 12 months from date of receipt.

Immunohistochemistry analysis using Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody, Clone J18. Tissue: Brain. Species: Mouse. Primary Antibody: Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody at 1:10000. Secondary Antibody: anti-rabbit HRP. C57/BL6 mice were injected with 5 ug sonicated mouse recombinant alpha synuclein PFFs at 8 weeks of age. Mice were unilaterally injected in the dorsal striatum (bregma AP + 0.2 mm, L +/1 2.0 mm, V - 3.0 mm) and sacrificed 30 days post-injection. (A) contralateral cortex. (B) ipsilateral cortex. (C) contralateral striatum. (D) ipsilateral striatum.

Immunocytochemistry/Immunofluorescence analysis using Rabbit Anti-Alpha Synuclein (pSer129) Monoclonal Antibody, Clone J18. Tissue: iPSC-derived neurons. Species: Human. Primary Antibody: Rabbit Anti-Alpha Synuclein (pSer129) Monoclonal Antibody at 1:1000 for O/N at 4°C. Secondary Antibody: Anti-Rabbit: A488 at 1:1000 for 1 hour at RT. Magnification: 40X. Nuclear stain: Hoechst- 20 min, RT (blue). Actin stain: Phalloidin-647- 20 min, RT (magenta). 4K cells per well. A) negative control; no fibrils added to well. B) 7 days after addition of active recombinant human pre-formed fibrils (Type 1).

Immunohistochemistry analysis using Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody, Clone J18. Tissue: Brain. Species: Mouse. Primary Antibody: Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody at 1:10000. Secondary Antibody: anti-rabbit HRP. C57/BL6 mice were injected with sonicated recombinant mouse alpha synuclein monomers or fibrils at 8 weeks of age. Mice were unilaterally injected in the dorsal striatum (bregma AP + 0.2 mm, L +/1 2.0 mm, V - 3.0 mm) and sacrificed 30 days post-injection. (A) 1.25 uL mouse alpha synuclein monomers. (B) 2.5 uL mouse alpha synuclein monomers. (C) 2.5 ug alpha synuclein PFFs. (D) 5 ug alpha synuclein PFFs. Inset: PBS (negative control).

Dot Blot analysis using Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody, Clone J18. Tissue: alpha synuclein peptide. Primary Antibody: Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody at 1:500 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-rabbit IgG:HRP at 1:4000 for 1 hour at RT 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 Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody, Clone J18. Lane 1: MW ladder. Lane 2: 0.5 ug human alpha synuclein monomer. Lane 3: 2 ug human alpha synuclein monomer. Lane 4: 0.5 ug human alpha synuclein PFFs. Lane 5: 2 ug human alpha synuclein PFFs. Lane 6: 15 ug human Parkinson's Disease brain lysate. Block: 5% BSA in TBST. Primary Antibody: Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody at 1:500 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-rabbit IgG:HRP at 1:4000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.

Western Blot analysis of Mouse Brain showing detection of Alpha Synuclein pSer129 protein using Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody, Clone J18. Lane 1: MW ladder. Lane 2: Mouse brain. Load: 15 ug. Block: 5% BSA in TBST. Primary Antibody: Rabbit Anti-Alpha Synuclein pSer129 Monoclonal Antibody at 1:500 for 2 hours at RT with shaking. Secondary Antibody: Goat anti-rabbit IgG:HRP at 1:4000 for 1 hour at RT with shaking. Color Development: Chemiluminescent for HRP (Moss) for 5 min in RT.

Immunocytochemistry / Immunofluorescence analysis of human iPSC-derived neurons treated with 2.5μg ATTO 594 labeled type I alpha-synuclein pre-formed fibrils for up to 14 days. Cells seeded at 8k cells per well. Green: mouse anti-alpha synuclein (pSer129) monoclonal antibody 1:5000; Red: alpha-synuclein PFFs; Pink: actin; Blue: Hoechst / DNA.