Recombinant
RabMAb

Anti-Alpha-synuclein filament antibody [MJFR-14-6-4-2] - Conformation-Specific (ab209538)

Overview

  • Product name
    Anti-Alpha-synuclein filament antibody [MJFR-14-6-4-2] - Conformation-Specific
    See all Alpha-synuclein filament primary antibodies
  • Description
    Rabbit monoclonal [MJFR-14-6-4-2] to Alpha-synuclein filament - Conformation-Specific
  • Tested applications
    Suitable for: IHC-FoFr, IHC-P, ICC/IF, Dot blotmore details
    Unsuitable for: WB
  • Species reactivity
    Reacts with: Mouse, Rat, Human
  • Immunogen

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

  • Positive control
    • Dot Blot: Recombinant alpha-synuclein filament untreated and treated with 70% formic acid; Recombinant alpha-synuclein monomer; ICC/IF: ReNcell VM differentiated in media containing cAMP and GDNF (without bFGF or EGF) and transduced with Ad5C01 viral vector encoding human alpha-synuclein filament.
  • General notes

    ab209538 is not suitable for WB or other denaturing conditions, as it is conformation-specific.

    This antibody is useful for studying Parkinson’s disease and other synucleinopathies including dementia with Lewy bodies and multiple system atrophy.

    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 ab209538 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-FoFr Use at an assay dependent concentration.
IHC-P Use at an assay dependent concentration.
ICC/IF 1/5000.
Dot blot Use a concentration of 0.0022 µg/ml.
  • Application notes
    Is unsuitable for WB.
  • 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
      • NACP antibody
      • Non-A beta component of AD amyloid antibody
      • Non-A4 component of amyloid precursor antibody
      • PARK1 antibody
      • PARK4 antibody
      • PD1 antibody
      • SNCA antibody
      • Synuclein alpha 140 antibody
      • Synuclein alpha antibody
      • SYUA_HUMAN antibody
      see all

    Images

    • ab209538 staining Alpha-synuclein filament in rat dorsal root ganglian (DRG) tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, cut into 20 micron slices and blocked with 2% BSA for 10 minutes at 21°C and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/2000 in TBS/BSA/azide) at 21°C for 2 hours. An Alexa Fluor® 555-conjugated Goat anti-rabbit polyclonal (1/300) was used as the secondary antibody.

      See Abreview

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized ReNcell VM (Human neural progenitor) cells labeling Alpha-synuclein filament with ab209538 at 1/5000 dilution, followed by Alexa Fluor® 647 Donkey anti-Rabbit IgG (H+L) secondary antibody at 1/400 dilution (red).

      Blocking buffer: 3% bovine serum albumin and 2% fetal bovine serum.

      ReNcell VM cells were differentiated in media containing cAMP and GDNF (without bFGF or EGF) and transduced with Ad5C01 viral vector encoding human alpha-synuclein filament (Left image). Right image show control vector cells.

      ab209538 specifically recognizes Alpha-synuclein filament.

      Images were reproduced courtesy of Charles River.

    • Ab209538 staining Alpha-synuclein filament in Human DLB brain tissue sections by Immunohistochemistry (Formalin/PFA fixed paraffin embedded sections). Tissue was fixed with paraformaldehyde and blocked with 2% BSA for 10 minutes at 21°C; antigen retrieval was by heat mediation in Citric acid. Samples were incubated with primary antibody (1/10,000 in TBS) for 2 hours at 21°C. A biotin conjugated anti-rabbit IgG Goat polyclonal was used as the secondary antibody at 1/300 dilution.

      See Abreview

    • Ab209538 staining Alpha-synuclein filament in transgenic α-Synuclein (A30P) mouse brain cells by IHC-FoFr (PFA perfusion fixed frozen sections). Tissue was fixed with paraformaldehyde and blocked with 10% serum for 1 hour at 24°C and permeabilized with 0.1m PBS with 3% Triton X. Samples were incubated with primary antibody (1/1000) for 24 hours at 4°C. An Alexa Fluor® 488 Donkey monoclonal was used as a secondary antibody at 1/3000 dilution.

      See Abreview

    • ab209538 staining Alpha-synuclein filament in mouse colon tissue sections by Immunohistochemistry (PFA perfusion fixed frozen sections). Tissue samples were fixed by perfusion with formaldehyde, cut into 20 micron slices, blocked with 2% BSA for 10 minutes at 21°C and antigen retrieval was by heat mediation in citrate buffer. The sample was incubated with primary antibody (1/2000 in TBS/BSA/azide) at 21°C for 2 hours. A biotin-conjugated goat anti-rabbit polyclonal (1/300) was used as the secondary antibody.

      See Abreview

    • Immunofluorescent analysis of 4% paraformaldehyde-fixed, 0.2% Triton X-100 permeabilized ReNcell VM (Human neural progenitor) cells labeling Alpha-synuclein filament with ab209538 at 1/5000 dilution, followed by Alexa Fluor® 647 Donkey anti-Rabbit IgG (H+L) secondary antibody at 1/400 dilution (red).

      Upper panel counterstain: Anti-aggregated alpha-synuclein antibody clone 5G4 at 1/400 dilution, followed by AlexaFluor®488 secondary detection (green).

      Lower panel counterstain: Anti-alpha/beta-synculein antibody at 1/200 dilution, followed by Alexa Fluor® 488 secondary detection (green).

      Blocking buffer: 3% bovine serum albumin and 2% fetal bovine serum.

      ReNcell VM cells were differentiated in media containing cAMP and GDNF (without bFGF or EGF) and transduced with Ad5C01 viral vector encoding human WT alpha-synuclein filament.

      ab209538 detects alpha-synuclein filaments, which are also recognized by anti-aggregated alpha-synuclein antibody clone 5G4 (upper panel). 

      Anti-alpha/beta-synculein antibody interacts with total alpha-synuclein in neurons, whereas ab209538 only recognizes the filamentous form (lower panel).    

      Images were reproduced courtesy of Charles River.

       

    • Dot blot analysis of alpha-synuclein filament labeled with ab209538 at 2.2 ng/ml.

      Lane 1: Recombinant alpha-synuclein filament treated with 70% formic acid.

      Lane 2: Recombinant alpha-synuclein filament treated with 70% formic acid.

      Lane 3: Untreated recombinant alpha-synuclein filament.

      Lane 4: Untreated recombinant alpha-synuclein filament.

      Alpha Synuclein filaments were generated using full length recombinant alpha-synuclein (aa1-140) by incubation in 20mM TRIS, pH 7.2 at 37 °C with agitation.

      Denaturation of filaments with 70% formic acid reduces antibody recognition by 30-100 fold, demonstrating conformation specificity.

      Data is provided by Professor Poul Henning Jensen, Aarhus University, Denmark.

    • Dot blot analysis of alpha synuclein labeled with ab209538 at 2 ng/ml.

      Lane 1: Recombinant alpha synuclein monomer.

      Lane 2: Recombinant alpha synuclein filament.

      Lane 3: Recombinant alpha synuclein monomer pre-absorbed with 100 µg/ml monomer alpha synuclein.

      Lane 4: Recombinant alpha synuclein filament pre-absorbed with 100 µg/ml monomer alpha-synuclein.

      Alpha-synuclein filaments were generated using full length recombinant alpha-synuclein (aa1-140) by incubation in 20mM TRIS, pH 7.2 at 37 °C with agitation.

      Pre-absorption with excess monomer demonstrates that the antibody recognizes soluble oligomeric species that spontaneously form in equilibrium with the monomer in solution.

      Data is provided by Professor Poul Henning Jensen, Aarhus University, Denmark.

       

    References

    ab209538 has not yet been referenced specifically in any publications.

    Customer reviews and Q&As

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (PFA perfusion fixed frozen sections)
    Sample
    Mouse Cell (Transgenic a-Synuclein (A30P) mouse brain)
    Permeabilization
    Yes - 0.1M PBS with 3% Triton X
    Specification
    Transgenic a-Synuclein (A30P) mouse brain
    Blocking step
    Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 24°C
    Fixative
    Paraformaldehyde
    Username

    Dr. Ruma Raha-Chowdhury

    Verified customer

    Submitted Sep 15 2016

    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Rat Tissue sections (DRG)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    DRG
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 29 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Mouse Tissue sections (Colon)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    Colon
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 29 2016

    Abcam guarantees this product to work in the species/application used in this Abreview.
    Application
    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
    Sample
    Human Tissue sections (DLB brain)
    Antigen retrieval step
    Heat mediated - Buffer/Enzyme Used: Citric acid
    Permeabilization
    No
    Specification
    DLB brain
    Blocking step
    BSA as blocking agent for 10 minute(s) · Concentration: 2% · Temperature: 21°C
    Fixative
    Formaldehyde
    Username

    Mr. Carl Hobbs

    Verified customer

    Submitted Feb 15 2016

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

    Sign up