Anti-alpha Synuclein (phospho S129) antibody [EP1536Y] (ab51253)

Overview

  • Product nameAnti-alpha Synuclein (phospho S129) antibody [EP1536Y]
    See all alpha Synuclein primary antibodies
  • Description
    Rabbit monoclonal [EP1536Y] to alpha Synuclein (phospho S129)
  • SpecificityThis 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. This antibody however showed high background in mouse and therefore we cannot guarantee it for use in IHC in mouse.
  • Tested applicationsSuitable for: WB, IHC-P, ICC/IFmore details
    Unsuitable for: Flow Cyt,IHC-Fr or IP
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

    Synthetic peptide (the amino acid sequence is considered to be commercially sensitive) corresponding to Human alpha Synuclein.
    (Peptide available as ab188826)

  • Positive control
    • IHC-P: FFPE Human Substantia Nigra normal and Parkinson.
  • General notes

    This product is a recombinant rabbit monoclonal antibody.

     

    Produced using Abcam’s RabMAb® technology. RabMAb® technology is covered by the following U.S. Patents, No. 5,675,063 and/or 7,429,487.

Properties

Applications

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
WB 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.
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.
ICC/IF Use at an assay dependent concentration.
  • Application notesIs unsuitable for Flow Cyt,IHC-Fr or IP.
  • Target

    • FunctionMay 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 specificityExpressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver. Concentrated in presynaptic nerve terminals.
    • Involvement in diseaseGenetic 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 similaritiesBelongs to the synuclein family.
    • DomainThe '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
      modifications
      Phosphorylated, 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 localizationCytoplasm, cytosol. Membrane. Nucleus. Cell junction, synapse. Secreted. Membrane-bound in dopaminergic neurons.
    • Information by UniProt
    • Database links
    • Alternative names
      • Alpha synuclein antibody
      • Alpha-synuclein antibody
      • Alpha-synuclein, isoform NACP140 antibody
      • alphaSYN antibody
      • MGC105443 antibody
      • MGC110988 antibody
      • MGC127560 antibody
      • MGC64356 antibody
      • NACP antibody
      • Non A beta component of AD amyloid antibody
      • Non A4 component of amyloid antibody
      • Non A4 component of amyloid precursor antibody
      • Non-A beta component of AD amyloid antibody
      • Non-A-beta component of alzheimers disease amyloid , precursor of antibody
      • Non-A4 component of amyloid precursor antibody
      • Non-A4 component of amyloid, precursor of antibody
      • OTTHUMP00000218549 antibody
      • OTTHUMP00000218551 antibody
      • OTTHUMP00000218552 antibody
      • OTTHUMP00000218553 antibody
      • OTTHUMP00000218554 antibody
      • PARK 1 antibody
      • PARK 4 antibody
      • PARK1 antibody
      • PARK4 antibody
      • Parkinson disease (autosomal dominant, Lewy body) 4 antibody
      • Parkinson disease familial 1 antibody
      • PD 1 antibody
      • PD1 antibody
      • SNCA antibody
      • Snca synuclein, alpha (non A4 component of amyloid precursor) antibody
      • SYN antibody
      • Synuclein alpha antibody
      • Synuclein alpha 140 antibody
      • Synuclein, alpha (non A4 component of amyloid precursor) antibody
      • SYUA_HUMAN antibody
      see all

    Anti-alpha Synuclein (phospho S129) antibody [EP1536Y] images

    • 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

      This image is courtesy of Professor Michael Goedert’s team (Sophie Morgan and Masami Masuda-Suzukake).

    • IHC image of alpha Synuclein (phospho S129) staining in Human Parkinson Substantia Nigra formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab51253, 10µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then 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

    • Dot blot analysis of alpha Synuclein (pS129) peptide (Lane 1), 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.

    • Direct ELISA antibody dose-response curve using ab51253. Antibody concentration of 0-5000 ng/mL. Antigen concentration of 1000 ng/mL. An alkaline phosphatase conjugated goat anti-rabbit IgG (1/1000) was used as the secondary antibody.

    References for Anti-alpha Synuclein (phospho S129) antibody [EP1536Y] (ab51253)

    This product has been referenced in:
    • Gispert S  et al. Potentiation of neurotoxicity in double-mutant mice with Pink1 ablation and A53T-SNCA overexpression. Hum Mol Genet 24:1061-76 (2015). WB, IHC-P, IHC-FrFl ; Mouse . Read more (PubMed: 25296918) »
    • Sasaki A  et al. Sensitive western blotting for detection of endogenous Ser129-phosphorylated a-synuclein in intracellular and extracellular spaces. Sci Rep 5:14211 (2015). WB ; Human . Read more (PubMed: 26381815) »

    See all 19 Publications for this product

    Product Wall

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Sample Human Cell lysate - whole cell (H4 cells)
    Loading amount 5 µg
    Specification H4 cells
    Gel Running Conditions Reduced Denaturing (4-20%Tris-Glycine gel)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C
    Username

    Abcam user community

    Verified customer

    Submitted Nov 03 2009

    Application Immunohistochemistry (PFA perfusion fixed frozen sections)
    Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 20°C
    Antigen retrieval step None
    Sample Human Tissue sections (PD patient brain sections, mesencephalon, 40 um)
    Specification PD patient brain sections, mesencephalon, 40 um
    Permeabilization Yes - 0.5% Triton-X 100
    Fixative Paraformaldehyde
    Username

    Miss. Oldriska Chutna

    Verified customer

    Submitted May 30 2014

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application Western blot
    Loading amount 20 µg
    Gel Running Conditions Reduced Denaturing (12% TGX gel Biorad)
    Sample Human Cell lysate - whole cell (neuronal SHY5)
    Specification neuronal SHY5
    Treatment okadaic acid for 6-8 hours 37 oC
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 23°C
    Username

    Ph.D.. Yien Kuo

    Verified customer

    Submitted Aug 30 2013

    Application Western blot
    Sample Mouse Tissue lysate - whole (brain)
    Gel Running Conditions Reduced Denaturing (12%)
    Loading amount 70 µg
    Treatment alpha synuclein
    Specification brain
    Blocking step Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C
    Username

    Abcam user community

    Verified customer

    Submitted Sep 23 2015

    1)Our initial testing with rat showed negative and that the antibody is not recommended for use in rat. However, based on sequence analysis, it shares 100% identity with rat.

    2)We previously tested rat brain, heart, kidney and spleen tissues l...

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    Thank you for contacting us.


    I am sorry to hear of the trouble which you have been experiencing when lot no. GR93844. We have not seen this issue with other lots of ab51253. This product is covered by our Abpromise guarantee. We are happ...

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    Merci pour votre réponse.

    Nous sommes très désolé de cette inconvenance. Veuillez croire que la qualité de nos produits est très important pour nous. J'ai transmis votre feedback aussi au laboratoire....

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    Merci beaucoup pour votre patience pendant que j'étais en correspondance avec le laboratoire.

    Effectivement, le laboratoire avait obtenu en signal faible avec ce lot à 1/500 et nous allons donc vous remplacer les deux dernier tu...

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    Vielen Dank für Ihren Anruf.

    Ich freue mich sehr, Ihnen gute Nachrichten überbringen zu können.

    Wir werdenab Ende der nächsten Woche ab51253 (Anti-alpha Synuclein (phospho S129) antibody [EP1536Y]) wieder anbiete...

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    Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"