Recombinant 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
Related conjugates and formulations
Overview
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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.
Mouse and rat species are recommended based on WB results, we do not guarantee IHC-FrFl and IHC-P for Mouse and rat.
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Tested applications
Suitable for: IHC-FrFl, WB, Dot blot, ELISA, IHC-Pmore details
Unsuitable for: Flow Cyt,IHC-Fr or IP -
Species reactivity
Reacts with: Mouse, Rat, Human
Predicted to work with: Cow, Pig, Chimpanzee, Macaque monkey, Gorilla, Orangutan, Spider monkey -
Immunogen
Synthetic peptide. This information is proprietary to Abcam and/or its suppliers.
(Peptide available asab188826) -
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. Mouse brain with Alzheimer’s disease tissue lysate with and without alkaline phosphatase incubation. IHC-Fl: Mouse brain tissue. ELISA: Alpha-synuclein (pS129) phospho peptide. Dot Blot: Human Alpha-synuclein (pS129) peptide.
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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
Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMAb® patents.
Properties
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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...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EP1536Y -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Immunizing Peptide (Blocking)
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Isotype control
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab51253 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Application | Abreviews | Notes |
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IHC-FrFl | (1) |
Use at an assay dependent concentration.
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WB | (4) |
1/1000 - 1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).Can be blocked with Alpha-synuclein (phospho S129) peptide (ab188826).
Good results have been obtained by treating the membrane with 0.4% PFA for 30 min at room temperature before blocking it with 5% milk. |
Dot blot |
Use at an assay dependent concentration.
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ELISA |
Use at an assay dependent concentration.
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IHC-P |
Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol.
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Notes |
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IHC-FrFl
Use at an assay dependent concentration. |
WB
1/1000 - 1/5000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).Can be blocked with Alpha-synuclein (phospho S129) peptide (ab188826). Good results have been obtained by treating the membrane with 0.4% PFA for 30 min at room temperature before blocking it with 5% milk. |
Dot blot
Use at an assay dependent concentration. |
ELISA
Use at an assay dependent concentration. |
IHC-P
Use a concentration of 5 - 10 µg/ml. Perform heat mediated antigen retrieval with citrate buffer pH 6 before commencing with IHC staining protocol. |
Target
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Function
May be involved in the regulation of dopamine release and transport. Induces fibrillization of microtubule-associated protein tau. Reduces neuronal responsiveness to various apoptotic stimuli, leading to a decreased caspase-3 activation. -
Tissue specificity
Expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver. Concentrated in presynaptic nerve terminals. -
Involvement in disease
Genetic alterations of SNCA resulting in aberrant polymerization into fibrils, are associated with several neurodegenerative diseases (synucleinopathies). SNCA fibrillar aggregates represent the major non A-beta component of Alzheimer disease amyloid plaque, and a major component of Lewy body inclusions. They are also found within Lewy body (LB)-like intraneuronal inclusions, glial inclusions and axonal spheroids in neurodegeneration with brain iron accumulation type 1.
Parkinson disease 1
Parkinson disease 4
Dementia Lewy body -
Sequence similarities
Belongs to the synuclein family. -
Domain
The 'non A-beta component of Alzheimer disease amyloid plaque' domain (NAC domain) is involved in fibrils formation. The middle hydrophobic region forms the core of the filaments. The C-terminus may regulate aggregation and determine the diameter of the filaments. -
Post-translational
modificationsPhosphorylated, predominantly on serine residues. Phosphorylation by CK1 appears to occur on residues distinct from the residue phosphorylated by other kinases. Phosphorylation of Ser-129 is selective and extensive in synucleinopathy lesions. In vitro, phosphorylation at Ser-129 promoted insoluble fibril formation. Phosphorylated on Tyr-125 by a PTK2B-dependent pathway upon osmotic stress.
Hallmark lesions of neurodegenerative synucleinopathies contain alpha-synuclein that is modified by nitration of tyrosine residues and possibly by dityrosine cross-linking to generated stable oligomers.
Ubiquitinated. The predominant conjugate is the diubiquitinated form.
Acetylation at Met-1 seems to be important for proper folding and native oligomeric structure. -
Cellular localization
Cytoplasm, cytosol. Membrane. Nucleus. Cell junction, synapse. Secreted. Membrane-bound in dopaminergic neurons. - Information by UniProt
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Database links
- Entrez Gene: 461375 Chimpanzee
- Entrez Gene: 101146366 Gorilla
- Entrez Gene: 6622 Human
- Entrez Gene: 20617 Mouse
- Entrez Gene: 100190854 Orangutan
- Entrez Gene: 641350 Pig
- Entrez Gene: 29219 Rat
- Omim: 163890 Human
see all -
Alternative names
- Alpha synuclein antibody
- Alpha-synuclein antibody
- Alpha-synuclein, isoform NACP140 antibody
see all
Images
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All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution
Lane 1 : In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of PLK2 (Polo-like kinase 2) but absence of ATP
Lane 2 : In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of ATP but absence of PLK2 (Polo-like kinase 2)
Lane 3 : In vitro kinase assay of Alpha Synuclein phosphorylation using His tagged human full length recombinant alpha-synuclein protein in the presence of PLK2 (Polo-like kinase 2) and ATP
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 17 kDa why is the actual band size different from the predicted?Blocking and Diluting buffer and concentration - 5% NFDM/TBST
Exposure time: 3 seconds
Lysates used here were prepared from 1% SDS hot method. Please refer to here.
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All lanes : Anti-Alpha-synuclein (phospho S129) antibody [EP1536Y] (ab51253) at 1/1000 dilution
Lane 1 : Mouse brain with Alzheimer’s disease tissue lysate
Lane 2 : Mouse brain with Alzheimer’s disease tissue lysate, membrane was incubated with alkaline phosphatase
Lysates/proteins at 20 µg per lane.
Secondary
All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/20000 dilution
Predicted band size: 14 kDa
Observed band size: 100,18 kDa why is the actual band size different from the predicted?
Exposure time: 140 secondsBlocking and diluting buffer and concentration: 5% NFDM/TBST
This blot was developed using a higher sensitivity ECL substrate.
Band around 100kda corresponds to αS oligomer (PMID: 27637918, PMID: 12597857)
Lysates used here were prepared using RIPA method. We recommend 1% SDS hot lysate method to reduce the detection of oligomers. Please refer here.
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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
Lysates used here were prepared from 1% SDS hot method. Please refer to here.
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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
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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
Lysates used here were prepared from 1% SDS hot method. Please refer to here.
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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.
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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 -
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.
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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.
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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).
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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 4oC and detected using a Biotinylated conjugated Anti-Rabbit monocloal antibody, 1/200.
Datasheets and documents
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SDS download
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Datasheet download
References (293)
ab51253 has been referenced in 293 publications.
- Tourville A et al. Modelling a-Synuclein Aggregation and Neurodegeneration with Fibril Seeds in Primary Cultures of Mouse Dopaminergic Neurons. Cells 11:N/A (2022). PubMed: 35626675
- Liu W et al. Characterization of a Novel Monoclonal Antibody for Serine-129 Phosphorylated α-Synuclein: A Potential Application for Clinical and Basic Research. Front Neurol 13:821792 (2022). PubMed: 35250825
- Reimer L et al. Low dose DMSO treatment induces oligomerization and accelerates aggregation of α-synuclein. Sci Rep 12:3737 (2022). PubMed: 35260646
- Stein CS et al. Modulation of miR-181 influences dopaminergic neuronal degeneration in a mouse model of Parkinson's disease. Mol Ther Nucleic Acids 28:1-15 (2022). PubMed: 35280925
- Fixemer S et al. Microglia phenotypes are associated with subregional patterns of concomitant tau, amyloid-β and α-synuclein pathologies in the hippocampus of patients with Alzheimer's disease and dementia with Lewy bodies. Acta Neuropathol Commun 10:36 (2022). PubMed: 35296366