Name: Anti-Alpha-synuclein antibody [EPR20535]
Brand: RabMAb
SKU: ab212184
Rating: 5 (2 reviews)

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Western blot - 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

Carrier Free

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

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...
Purity

Protein A purified
Clonality

Monoclonal
Clone number

EPR20535
Isotype

IgG
Research areas
- Neuroscience
- Diseases
- Neuroscience
- Processes

Alternative Versions
- Anti-Alpha-synuclein antibody [EPR20535] - BSA and Azide free (ab225866)
Compatible Secondaries
Isotype control
- Rabbit IgG, monoclonal [EPR25A] - Isotype Control (ab172730)
Recombinant Protein
- Recombinant Human Alpha-synuclein protein aggregate (Active) (ab218819)
Related Products
- VeriBlot for IP Detection Reagent (HRP) (ab131366)
- Antigen Retrieval Buffer (100X Tris-EDTA Buffer, pH 9.0) (ab93684)

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
IHC-P		1/16000. Perform heat mediated antigen retrieval with Tris/EDTA buffer pH 9.0 before commencing with IHC staining protocol.
WB	(1)	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.

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.

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.
Target information above from: UniProt accession P37840 The UniProt Consortium
The Universal Protein Resource (UniProt) in 2010
Nucleic Acids Res. 38:D142-D148 (2010) .

Information by UniProt
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
- Unigene: 17484 Mouse
- Unigene: 1827 Rat
see all
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

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

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.
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, 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^e® 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.
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)

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

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).
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)

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.
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)

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)

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

Immunoprecipitation protocols
Immunohistochemistry protocols
Western blot protocols

Click here to view the general protocols

SDS download

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Datasheet download

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Publishing research using ab212184? Please let us know so that we can cite the reference in this datasheet.

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

View all Publications for this product

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1-2 of 2 Abreviews or Q&A

Application

Western blot

Sample

Mouse Tissue lysate - whole (mouse brain)

Gel Running Conditions

Reduced Denaturing (4-20% gel)

Loading amount

15 µg

Specification

mouse brain

Blocking step

Intercept® (TBS) Blocking Buffer as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 100% · Temperature: 24°C

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Jie Zhang

Verified customer

Submitted Apr 09 2021

Application

Immunohistochemistry free floating

Sample

Mouse Tissue sections (Caudate Putamen)

Specification

Caudate Putamen

The reviewer received a reward from Abcam’s Loyalty Program in thanks for submitting this Abreview and for helping the scientific community make better-informed decisions.

Abcam user community

Verified customer

Submitted Apr 29 2019

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Key features and details

Related conjugates and formulations

Overview

Product name

Description

Host species

Tested applications

Species reactivity

Immunogen

Positive control

General notes

Properties

Form

Storage instructions

Storage buffer

Purity

Clonality

Clone number

Isotype

Research areas

Associated products

Alternative Versions

Compatible Secondaries

Isotype control

Recombinant Protein

Related Products

Applications

The Abpromise guarantee

Target

Function

Tissue specificity

Involvement in disease

Sequence similarities

Domain

Post-translationalmodifications

Cellular localization

Database links

Alternative names

Images

Protocols

Datasheets and documents

SDS download

Datasheet download

Certificate of Compliance

References (13)

Customer reviews and Q&As

Western blot abreview for Anti-Alpha-synuclein antibody [EPR20535]

Immunohistochemistry free floating abreview for Anti-Alpha-synuclein antibody [EPR20535]

Post-translational
modifications