Overview

  • Product name

    Anti-Alpha-synuclein antibody [EPR20535]
    See all Alpha-synuclein primary antibodies
  • Description

    Rabbit monoclonal [EPR20535] to Alpha-synuclein
  • Host species

    Rabbit
  • Tested applications

    Suitable for: IHC-P, WB, IHC-Fr, IPmore details
  • Species reactivity

    Reacts with: Mouse, Rat, Human
  • Immunogen

    Recombinant full length protein within Human Alpha-synuclein aa 1 to the C-terminus. The exact sequence is proprietary.
    Database link: P37840

  • Positive control

    • WB: Human Alpha-synuclein recombinant protein; Human cerebellum and brain lysates; Mouse and rat brain lysates. IHC-P: Human cerebral cortex and glioma tissues; Mouse and rat cerebral cortex tissues. IHC-Fr: Mouse hippocampus tissue. IP: Mouse brain lysate.
  • General notes

    Our RabMAb® technology is a patented hybridoma-based technology for making rabbit monoclonal antibodies. For details on our patents, please refer to RabMab® patents.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

Our Abpromise guarantee covers the use of ab212184 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
IHC-P 1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB 1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).
IHC-Fr 1/100.

Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20).

IP 1/30.

Target

  • 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
    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 localization

    Cytoplasm, 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
    • SNCA antibody
    • Snca synuclein 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

Images

  • Lane 1: Wild type HAP1 whole cell lysate (20 µg)
    Lane 2: SNCA (alpha Synuclein) knockout HAP1 whole cell lysate (20 µg)
    Lane 3: Human brain whole tissue lysate (20 µg)
    Lane 4: SH-SY5Y whole cell lysate (20 µg)

    Lanes 1 - 4: Merged signal (red and green). Green - ab212184 observed at 14 kDa. Red - loading control, ab9484, observed at 37 kDa.

    ab212184 was shown to specifically react with SNCA (alpha Synuclein) in wild type cells as signal was lost in SNCA (alpha Synuclein) knockout cells. Wild-type and SNCA (alpha Synuclein) knockout samples were subjected to SDS-PAGE. ab212184 and ab9484 (Mouse anti-GAPDH loading control) were incubated overnight at 4°C at a 1/1000 dilution and 1/20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDy 800CW) preabsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

  • Immunohistochemical analysis of paraffin-embedded human cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Cytoplasmic staining on human cerebral cortex [PMID: 22112368].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized frozen mouse hippocampus tissue labeling Alpha-synuclein with ab212184 at 1/100 dilution, followed by Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) secondary antibody at 1/1000 dilution (green).

    Cytoplasmic staining on mouse hippocampus (PMID: 22112368).

    The nuclear counterstain is DAPI (blue).

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat anti-rabbit IgG (Alexa Fluor® 488) (ab150077) at 1/1000 dilution.

  • All lanes : Anti-Alpha-synuclein antibody [EPR20535] (ab212184) at 1/1000 dilution

    Lane 1 : Human cerebellum lysate
    Lane 2 : Human brain lysate
    Lane 3 : Mouse brain lysate
    Lane 4 : Rat brain lysate

    Lysates/proteins at 20 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 5 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    The observed molecular weight is consistent with the literature (PMID: 11739566).

  • Immunohistochemical analysis of paraffin-embedded human glioma tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Cytoplasmic staining on human glioma [PMID: 22112368].
    Counter stained with Hematoxylin.
     
    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
  • All lanes : Anti-Alpha-synuclein antibody [EPR20535] (ab212184) at 1/1000 dilution

    Lane 1 : Human Alpha-synuclein recombinant protein
    Lane 2 : Human Beta-synuclein recombinant protein

    Lysates/proteins at 0.01 µg per lane.

    Secondary
    All lanes : Goat Anti-Rabbit IgG H&L (HRP) (ab97051) at 1/100000 dilution

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


    Exposure time: 8 seconds


    Blocking/Dilution buffer: 5% NFDM/TBST.

    Human Alpha-synuclein recombinant protein contain aa1-140 with His-tag. Human Beta-synuclein recombinant protein contain aa1-134 with His-tag.

  • Immunohistochemical analysis of paraffin-embedded mouse cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Cytoplasmic staining on mouse cerebral cortex [PMID: 22112368].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded rat cerebral cortex tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Cytoplasmic staining on rat cerebral cortex [PMID: 22112368].

    Counter stained with Hematoxylin.

    Secondary antibody only control: Used PBS instead of primary antibody, secondary antibody is Goat Anti-Rabbit IgG H&L (HRP) Ready to use.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Immunohistochemical analysis of paraffin-embedded human kidney tissue labeling Alpha-synuclein with ab212184 at 1/16000 dilution, followed by Goat Anti-Rabbit IgG H&L (HRP) Ready to use.
     
    Negative control: No staining on human kidney. [PMID: 14997013].

    Counter stained with Hematoxylin.

    Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.

  • Alpha-synuclein was immunoprecipitated from 0.35 mg of mouse brain lysate with ab212184 at 1/30 dilution.

    Western blot was performed from the immunoprecipitate using ab212184 at 1/1000 dilution.

    VeriBlot for IP Detection Reagent (HRP) (ab131366), was used for detection at 1/1000 dilution

    Lane 1: Mouse brainlysate 10ug (Input).

    Lane 2: ab212184 IP in mouse brain lysate.

    Lane 3: Rabbit monoclonal IgG (ab172730) instead of ab212184 in mouse brain lysate.

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

    Exposure time: 1 second.

References

This product has been referenced in:

  • Canerina-Amaro A  et al. Differential Aggregation and Phosphorylation of Alpha Synuclein in Membrane Compartments Associated With Parkinson Disease. Front Neurosci 13:382 (2019). Read more (PubMed: 31068782) »
  • Schaser AJ  et al. Alpha-synuclein is a DNA binding protein that modulates DNA repair with implications for Lewy body disorders. Sci Rep 9:10919 (2019). Read more (PubMed: 31358782) »
See all 2 Publications for this product

Customer reviews and Q&As

Filter by Application

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Application
Immunohistochemistry free floating
Sample
Mouse Tissue sections (Caudate Putamen)
Specification
Caudate Putamen

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Verified customer

Submitted Apr 29 2019

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