Antibodies, Proteins, Kits and Reagents for Life Science

388 646 ₸ Product size

100 µl 388 646 ₸ 10 µl 154 118 ₸

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Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Key features and details

Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
Rabbit monoclonal [EP1536Y] to Alpha-synuclein (phospho S129)
Suitable for: IHC-FrFl, WB, Dot blot, ELISA, IHC-P
Reacts with: Mouse, Rat, Human

Product name

Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y]
See all Alpha-synuclein primary antibodies
Description

Rabbit monoclonal [EP1536Y] to Alpha-synuclein (phospho S129)
Host species

Rabbit
Specificity

This antibody only detects alpha synuclein phosphorylated on Ser129. IHC-P: This antibody showed no staining in human hippocampus normal brain and showed staining in Parkinson's brain as expected.

Tested Applications & Species

Application	Species
Dot	Human
IHC-FrFl	Mouse
IHC-P	Human
WB	Mouse Rat

See all applications and species data

Immunogen

Synthetic peptide within Human Alpha-synuclein aa 100 to the C-terminus. The exact sequence is proprietary.
Database link: P37840
(Peptide available as ab188826)
Positive control
- IHC-P: Human Parkinson Substantia Nigra tissue. WB: Sarkosyl-insoluble brain extract from mice transgenic for PrPA53T alpha-synuclein; Recombinant alpha-synuclein phosphorylated at S129; Rat and mouse brain lysates. IHC-Fl: Mouse brain tissue. ELISA: Alpha-synuclein (pS129) phospho peptide. Dot Blot: Human Alpha-synuclein (pS129) peptide.
General notes
This product is a recombinant monoclonal antibody, which offers several advantages including:
- - High batch-to-batch consistency and reproducibility
- - Improved sensitivity and specificity
- - Long-term security of supply
- - Animal-free production
For more information see here.
Our RabMAb^® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb^® patents.

Form

Liquid
Storage instructions

Shipped at 4°C. Upon delivery aliquot and store at -20°C. Avoid freeze / thaw cycles.
Storage buffer

pH: 7.20
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.5% BSA
Concentration information loading...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EP1536Y
Isotype

IgG
Research areas

Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Rat brain lysates, the membrane was incubated with alkaline phosphatase.
Lane 2 : Rat brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 14 kDa
Observed band size: 18 kDa
why is the actual band size different from the predicted?

Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 20 seconds
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

IHC image of alpha Synuclein (phospho S129) staining Human Parkinson Substantia Nigra tissue section*, previously antigen was retrieved by heat mediated with citrate buffer pH 6, fixed in formalin and embedded in paraffin. This section was incubated with ab51253 at 10 µg/mL for 15 mins at room temperature and detected using an HRP conjugated compact polymer system, performed on a Leica Bond™ system using the standard protocol F. DAB was used as the chromogen. Counterstained with haematoxylin and mounted with DPX. The inset negative control image is taken from an identical assay performed on Human normal Substantia Nigra.

For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.

*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Mouse brain lysates, the membrane was incubated with alkaline phosphatase
Lane 2 : Mouse brain lysates

Lysates/proteins at 15 µg per lane.

Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/2000 dilution

Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?

Blocking/Diluting buffer and concentration: 5% NFDM/TBST.

Exposure time: 40 seconds
Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) This image is taken from Machado de Oliveira. R et al. PLoS Biol. 2017 Mar; 15(3): e2000374. S7a doi: 10.1371/journal.pbio.2000374 Reproduced under the Creative Commons license http://creativecommons.org/licenses/by/4.0/.

Expression of WT aSyn in the SN is toxic over time. Mouse brain sections immunostained for TH (red panels) and aSyn (using ab51253) (green panels) 1, 2 and 3 weeks after injection with vectors encoding for EGFP or WT aSyn. Scale bar for isolated channels 200 μm and for merged channels 100 μm.
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) This image is courtesy of Professor Michael Goedert’s team (Sophie Morgan and Masami Masuda-Suzukake).

All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution

Lane 1 : Sarkosyl-insoluble brain extract from mice transgenic for PrPA53T alpha-synuclein (line M83)
Lane 2 : Recombinant alpha-synuclein phosphorylated at S129, 3ng

Predicted band size: 14 kDa
ELISA - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Direct ELISA antibody dose-response curve using ab51253.

Antibody concentration of 0-5000 ng/mL.

Antigen (Alpha-synuclein (pS129) phospho peptide and Alpha-synuclein non-phospho peptide) concentration of 1000 ng/mL.

An alkaline phosphatase conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.
Dot Blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Dot blot analysis of human alpha Synuclein (pS129) peptide (Lane 1), huamn alpha Synuclein (unmodified) peptide (Lane 2) labelling alpha Synuclein (pS129) with ab51253 at a dilution of 1/1000. Peroxidase conjugated goat anti-rabbit IgG (H+L) was used as the secondary antibody at a dilution of 1/2500.

Blocking and dilution buffer: 5% NFDM/TBST.

Exposure time: 3 minutes.
Western blot - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) This image is taken from Migdalska-Richards A. et al. Ann Neurol. 2016 Nov; 80(5): 766–775. Fig 5 doi: 10.1002/ana.24790

α-Synuclein (α-syn) and S129-phosphorylated α-synuclein protein levels in SNCA/SNCA mouse brains after 12 days of treatment with 4mM ambroxol (Amb). (A) Western blotting for α-synuclein (using ab1903) and serine 129 (S129)-phosphorylated α-synuclein protein (using ab51253) in the brainstem (example blots shown).
Immunohistochemistry - Free Floating - Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) This image is courtesy of an abreview by Sophie Morgan

IHC image of alpha Synuclein (phospho S129) staining in free floating tgM83^+/- or tgM83^+/+ mouse brain tissue. The section was incubated with ab51253, 1/5000, for 15 hours at 4^oC and detected using a Biotinylated conjugated Anti-Rabbit monocloal antibody, 1/200.
See Abreview
Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253)

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