Skip to main content

Anti-Alpha-synuclein antibody [EPR20535] ab212184 is a rabbit monoclonal antibody that is used in Alpha-synuclein western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with SNCA knockout cell line validation
- Antibody clone EPR20535 is cited in over 30 publications


Images

Western blot - Anti-Alpha-synuclein antibody [EPR20535] (AB212184), expandable thumbnail
  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (AB212184), expandable thumbnail
  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (AB212184), expandable thumbnail
  • Immunohistochemistry (Frozen sections) - Anti-Alpha-synuclein antibody [EPR20535] (AB212184), expandable thumbnail
  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (AB212184), expandable thumbnail

Publications

Key facts

Isotype

IgG

Host species

Rabbit

Storage buffer

pH: 7.2 - 7.4
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol (glycerin, glycerine), 0.05% BSA

Form

Liquid

Clonality

Monoclonal

Immunogen

  • The exact immunogen used to generate this antibody is proprietary information.

Reactivity data

Select an application
Product promiseTestedExpectedPredictedNot recommended
WBIHC-PIHC-FrIP
Human
Tested
Tested
Expected
Expected
Mouse
Tested
Tested
Tested
Tested
Rat
Tested
Tested
Expected
Expected
Recombinant full length protein
Expected
Expected
Expected
Expected

Tested
Tested

Species

Rat

Dilution info

1/1000

Notes

-

Species

Mouse

Dilution info

1/1000

Notes

-

Species

Human

Dilution info

1/1000

Notes

-

Expected
Expected

Species

Recombinant full length protein

Dilution info

1/1000

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/16000

Notes

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

Species

Rat

Dilution info

1/16000

Notes

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

Species

Human

Dilution info

1/16000

Notes

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

Expected
Expected

Species

Recombinant full length protein

Dilution info

1/16000

Notes

-

Tested
Tested

Species

Mouse

Dilution info

1/100

Notes

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

Expected
Expected

Species

Rat

Dilution info

-

Notes

-

Species

Human

Dilution info

-

Notes

-

Species

Recombinant full length protein

Dilution info

1/100

Notes

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

Tested
Tested

Species

Mouse

Dilution info

1/30

Notes

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

Expected
Expected

Species

Rat

Dilution info

-

Notes

-

Species

Human

Dilution info

-

Notes

-

Species

Recombinant full length protein

Dilution info

1/30

Notes

-

Associated Products

Select an associated product type

15 products for Alternative Product

Target data

Function

Neuronal protein that plays several roles in synaptic activity such as regulation of synaptic vesicle trafficking and subsequent neurotransmitter release (PubMed:20798282, PubMed:26442590, PubMed:28288128, PubMed:30404828). Participates as a monomer in synaptic vesicle exocytosis by enhancing vesicle priming, fusion and dilation of exocytotic fusion pores (PubMed:28288128, PubMed:30404828). Mechanistically, acts by increasing local Ca(2+) release from microdomains which is essential for the enhancement of ATP-induced exocytosis (PubMed:30404828). Acts also as a molecular chaperone in its multimeric membrane-bound state, assisting in the folding of synaptic fusion components called SNAREs (Soluble NSF Attachment Protein REceptors) at presynaptic plasma membrane in conjunction with cysteine string protein-alpha/DNAJC5 (PubMed:20798282). This chaperone activity is important to sustain normal SNARE-complex assembly during aging (PubMed:20798282). Also plays a role in the regulation of the dopamine neurotransmission by associating with the dopamine transporter (DAT1) and thereby modulating its activity (PubMed:26442590).

Alternative names

Recommended products

Anti-Alpha-synuclein antibody [EPR20535] ab212184 is a rabbit monoclonal antibody that is used in Alpha-synuclein western blotting and IHC. Suitable for human, mouse and rat samples.

- Recombinant format for unrivaled batch-batch consistency: no need for same-lot requests
- Specificity confirmed with SNCA knockout cell line validation
- Antibody clone EPR20535 is cited in over 30 publications

Key facts

Isotype

IgG

Form

Liquid

Clonality

Monoclonal

Immunogen
  • The exact immunogen used to generate this antibody is proprietary information.
Clone number

EPR20535

Purification technique

Affinity purification Protein A

Concentration
Loading...

Storage

Shipped at conditions

Blue Ice

Appropriate short-term storage duration

1-2 weeks

Appropriate short-term storage conditions

+4°C

Appropriate long-term storage conditions

-20°C

Aliquoting information

Upon delivery aliquot

Storage information

Avoid freeze / thaw cycle

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 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 batch production

For more information, read more on recombinant antibodies.

Supplementary info

This supplementary information is collated from multiple sources and compiled automatically.

Activity summary

Alpha-synuclein often referred to by alternate names such as SNCA is a protein of around 14 kDa mass. It mainly expresses in the brain particularly in presynaptic nerve terminals. This protein functions mechanically by stabilizing synaptic vesicles and maintaining synaptic function. It exists both in soluble monomer forms and as aggregates in protein filaments. Antibodies like 4D6 and EP1536Y target monomer forms of protein for more detailed studies.

Biological function summary

The alpha-synuclein protein plays critical roles in neuronal activity. It contributes to neurotransmitter release regulation by acting in the formation and plasticity of the presynaptic neuronal network. Alpha-synuclein doesn't usually form parts of large protein complexes but it may associate transiently with membranes and vesicular structures. The protein's monomer form has also been observed in alpha lines and related neuronal processes operating alongside various cellular functions.

Pathways

Synaptic vesicle trafficking and dopamine neurotransmitter release are significant areas involving the alpha-synuclein protein. In these pathways alpha-synuclein interacts with other proteins like synaptophysin and protein monomer monomerizations are intrinsic to these processes. Altered function or aggregation of alpha-synuclein disrupts these pathways influencing broader neurological functions.

Associated diseases and disorders

Alterations or accumulations of alpha-synuclein are strongly linked to Parkinson's disease and Lewy body dementia. In these conditions alpha-synuclein forms abnormal protein filaments known as Lewy bodies within neurons. These formations disrupt cellular processes and neuron health. Synucleinopathies such as these show connections with proteins like parkin and DJ-1 which also have key roles in these neurodegenerative diseases.

Product promise

We are dedicated to supporting your work with high quality reagents and we are here for you every step of the way should you need us.

In the unlikely event of one of our products not working as expected, you are covered by our product promise.

Full details and terms and conditions can be found here:
Terms & Conditions.

13 product images

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    False colour image of Western blot: Anti-Alpha-synuclein antibody [EPR20535] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (Anti-GAPDH antibody [6C5] - Loading Control ab8245) loading control staining at 1/20000 dilution, shown in red. In Western blot, ab212184 was shown to bind specifically to Alpha-synuclein. A band was observed at 16 kDa in wild-type U-87 MG cell lysates with no signal observed at this size in SNCA knockout cell line Human SNCA (Alpha-synuclein) knockout U-87 MG cell line ab282333 (knockout cell lysate Human SNCA (Alpha-synuclein) knockout U-87 MG cell lysate ab283006). To generate this image, wild-type and SNCA knockout U-87 MG cell lysates were analysed. First, samples were run on an SDS-PAGE gel then transferred onto a nitrocellulose membrane. Membranes were blocked in 3% milk in TBS-0.1 % Tween® 20 (TBS-T) before incubation with primary antibodies overnight at 4°C. Blots were washed four times in TBS-T, incubated with secondary antibodies for 1 h at room temperature, washed again four times then imaged. Secondary antibodies used were Goat anti-Rabbit IgG H&L (IRDye® 800CW) preabsorbed (Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776) at 1/20000 dilution.

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

    Lane 1: Wild-type U-87 MG cell lysate at 20 µg

    Lane 2: SNCA knockout U-87 MG cell lysate at 20 µg

    Performed under reducing conditions.

    Predicted band size: 14 kDa

    Observed band size: 16 kDa

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    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, Anti-GAPDH antibody [mAbcam 9484] - 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 Anti-GAPDH antibody [mAbcam 9484] - Loading Control 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 Goat anti-Rabbit IgG H&L (IRDye® 800CW) preadsorbed ab216773 and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed Goat anti-Mouse IgG H&L (IRDye® 680RD) preadsorbed ab216776 secondary antibodies at 1/20000 dilution for 1 hour at room temperature before imaging.

    All lanes: Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    Predicted band size: 14 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)


    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.

  • Immunohistochemistry (Frozen sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Frozen sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    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) (Goat Anti-Rabbit IgG H&L (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) (Goat Anti-Rabbit IgG H&L (Alexa Fluor® 488) ab150077) at 1/1000 dilution.

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    Blocking/Dilution buffer: 5% NFDM/TBST.

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

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

    Lane 1: Human cerebellum lysate at 20 µg

    Lane 2: Human brain lysate at 20 µg

    Lane 3: Mouse brain lysate at 20 µg

    Lane 4: Rat brain lysate at 20 µg

    Secondary

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

    Predicted band size: 14 kDa

    Observed band size: 18 kDa

    Exposure time: 5s

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)


    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.

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    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.

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

    Lane 1: Human Alpha-synuclein recombinant protein at 0.01 µg

    Lane 2: Human Beta-synuclein recombinant protein at 0.01 µg

    Secondary

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

    Predicted band size: 14 kDa

    Observed band size: 18 kDa

    Exposure time: 8s

  • Immunoprecipitation - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunoprecipitation - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    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) (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 (Rabbit IgG, monoclonal [EPR25A] - Isotype Control ab172730) instead of ab212184 in mouse brain lysate.

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

    Exposure time: 1 second.

    All lanes: Immunoprecipitation - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    Predicted band size: 14 kDa

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)


    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)


    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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)


    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.

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    In Western blot, Anti-Alpha-synuclein antibody [EPR20535] (ab212184) staining at 1/1000 dilution.

    In Western blot, Anti-6X His tag® antibody [EPR20547] - ChIP Grade (Anti-6X His tag® antibody [EPR20547] - ChIP Grade ab213204) staining at 1/5000 dilution.

    All lanes: Western blot - Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] (Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] ab317366) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) transfected with humanTyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein (WT) expression vector containing a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate at 4 µg

    Lane 2: 293T transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein (mutant Y39) protein expression vector containing a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate at 4 µg

    Secondary

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

    Observed band size: 25 kDa

    Exposure time: 10s

  • Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184), expandable thumbnail

    Western blot - Anti-Alpha-synuclein antibody [EPR20535] (ab212184)

    In Western blot, Anti-Alpha-synuclein antibody [EPR20535] (ab212184) staining at 1/1000 dilution.

    All lanes: Western blot - Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] (Anti-Alpha-synuclein (phospho Y39) antibody [EPR25118-5] ab317366) at 1/1000 dilution

    Lane 1: 293T (human embryonic kidney epithelial cell) transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein expression vector containi a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate (untreated membrane) at 4 µg

    Lane 2: 293T transfected with human Tyrosine-protein kinase ABL1 expression vector and a human Alpha-synuclein protein expression vector containi a myc-His-tag® treated with 1 mM pervanadate for 30 minutes whole cell lysate (alkaline phosphatase treated membrane) at 4 µg

    Secondary

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

    Observed band size: 25 kDa

    Exposure time: 8s

Downloads

Product protocols

For this product, it's our understanding that no specific protocols are required. You can:

Please note: All products are 'FOR RESEARCH USE ONLY. NOT FOR USE IN DIAGNOSTIC OR THERAPEUTIC PROCEDURES'.

For licensing inquiries, please contact partnerships@abcam.com