Recombinant Anti-Alpha-synuclein antibody [EPR20535] (ab212184)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [EPR20535] to Alpha-synuclein
- Suitable for: IHC-P, WB, IHC-Fr, IP
- Knockout validated
- Reacts with: Mouse, Rat, Human, Recombinant fragment
Related conjugates and formulations
Overview
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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, Recombinant fragment -
Immunogen
Recombinant full length protein. This information is proprietary to Abcam and/or its suppliers.
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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.
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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. Store at +4°C short term (1-2 weeks). Upon delivery aliquot. Store at -20°C long term. Avoid freeze / thaw cycle. -
Storage buffer
pH: 7.2
Preservative: 0.01% Sodium azide
Constituents: 59% PBS, 40% Glycerol, 0.05% BSA -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
EPR20535 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Compatible Secondaries
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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 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 |
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IHC-P |
1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
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WB | (1) |
1/1000. Detects a band of approximately 18 kDa (predicted molecular weight: 14 kDa).
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IHC-Fr |
1/100.
Antigen retrieval: Heated citrate solution (10mM citrate pH 6.0 + 0.05% Tween-20). |
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IP |
1/30.
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Notes |
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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
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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: 6622 Human
- Entrez Gene: 20617 Mouse
- Entrez Gene: 29219 Rat
- Omim: 163890 Human
- SwissProt: P37840 Human
- SwissProt: O55042 Mouse
- SwissProt: P37377 Rat
- Unigene: 21374 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 antibody [EPR20535] (ab212184) at 1/1000 dilution
Lane 1 : Wild-type U-87 MG cell lysate
Lane 2 : SNCA knockout U-87 MG cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 16 kDa why is the actual band size different from the predicted?False colour image of Western blot: Anti-Alpha-synuclein antibody [EPR20535] staining at 1/1000 dilution, shown in green; Mouse anti-GAPDH antibody [6C5] (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 ab282333 (knockout 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 (ab216773) and Goat anti-Mouse IgG H&L (IRDye® 680RD) preabsorbed (ab216776) at 1/20000 dilution.
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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 (IRDye® 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.
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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.
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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.
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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 secondsBlocking/Dilution buffer: 5% NFDM/TBST.
The observed molecular weight is consistent with the literature (PMID: 11739566).
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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.
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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 secondsBlocking/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.
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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.
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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.
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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.
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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.
Protocols
Datasheets and documents
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SDS download
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Datasheet download
Certificate of Compliance
References (13)
ab212184 has been referenced in 13 publications.
- Patterson JR et al. Beta2-adrenoreceptor agonist clenbuterol produces transient decreases in alpha-synuclein mRNA but no long-term reduction in protein. NPJ Parkinsons Dis 8:61 (2022). PubMed: 35610264
- Yan YQ et al. Different patterns of exosomal α-synuclein between Parkinson's disease and probable rapid eye movement sleep behavior disorder. Eur J Neurol 29:3590-3599 (2022). PubMed: 36047985
- Azam S et al. α-Synuclein upregulates bim-mediated apoptosis by negatively regulating endogenous GCN5. Aging (Albany NY) 14:8292-8301 (2022). PubMed: 36309909
- Yang X et al. Time-dependent alterations in the rat nigrostriatal system after intrastriatal injection of fibrils formed by α-Syn and tau fragments. Front Aging Neurosci 14:1049418 (2022). PubMed: 36518823
- Ramalingam M et al. Neural-Induced Human Adipose Tissue-Derived Stem Cells Conditioned Medium Ameliorates Rotenone-Induced Toxicity in SH-SY5Y Cells. Int J Mol Sci 22:N/A (2021). PubMed: 33652595