Recombinant Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
Key features and details
- Produced recombinantly (animal-free) for high batch-to-batch consistency and long term security of supply
- Rabbit monoclonal [MJFR1] to Alpha-synuclein
- Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IF
- Knockout validated
- Reacts with: Human
Related conjugates and formulations
Overview
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Product name
Anti-Alpha-synuclein antibody [MJFR1]
See all Alpha-synuclein primary antibodies -
Description
Rabbit monoclonal [MJFR1] to Alpha-synuclein -
Host species
Rabbit -
Tested applications
Suitable for: Flow Cyt (Intra), WB, IHC-P, IP, ICC/IFmore details -
Species reactivity
Reacts with: Human -
Immunogen
Recombinant full length protein within Human Alpha-synuclein aa 1 to the C-terminus. The exact sequence is proprietary.
Database link: P37840 -
Epitope
The epitope was mapped to amino acids 118-123 (VDPDNE). -
Positive control
- WB: Recombinant Human Alpha-synuclein protein (ab51189), HAP1 and HEK293-T cell lysate; Human brain whole cell lysate; Human brain tissue lysate. IHC-P: FFPE Human Normal Cerebral Cortex; FFPE Human Normal and Parkinson Substantia Nigra tissue. Flow Cyt (intra): Hap1 cells. ICC/IF: Hap1 and U87-MG cells.
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General notes
Alpha-Synuclein is expressed predominantly in the brain, where it is concentrated in presynaptic nerve terminals. The deposition of the abundant presynaptic brain protein alpha-synuclein as fibrillary aggregates in neurons or glial cells is a hallmark lesion in a subset of neurodegenerative disorders. These disorders include Parkinson's disease (PD), dementia with Lewy bodies (DLB) and multiple system atrophy, collectively referred to as synucleinopathies. Parkinson's disease (PD) is a common neurodegenerative disorder characterized by the progressive accumulation in selected neurons of protein inclusions containing alpha-synuclein and ubiquitin.
This antibody was developed with support from The Michael J. Fox Foundation, in collaboration with the laboratory of Dr. Michael Schlossmacher (University of Ottawa).
Abcam recommended secondaries - Goat Anti-Rabbit HRP (ab205718) and Goat Anti-Rabbit Alexa Fluor® 488 (ab150077).
See other anti-rabbit secondary antibodies that can be used with this antibody.
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.
Rat: We have preliminary internal testing data to indicate this antibody may not react with this species. Please contact us for more information.
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. Avoid freeze / thaw cycle. -
Storage buffer
Preservative: 0.01% Sodium azide
Constituents: 40% Glycerol, 0.05% BSA, 59% PBS -
Concentration information loading...
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Purity
Protein A purified -
Clonality
Monoclonal -
Clone number
MJFR1 -
Isotype
IgG -
Research areas
Associated products
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Alternative Versions
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Conjugation kits
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Isotype control
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KO cell lines
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KO cell lysates
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Positive Controls
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Recombinant Protein
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Related Products
Applications
The Abpromise guarantee
Our Abpromise guarantee covers the use of ab138501 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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Flow Cyt (Intra) |
1/200.
For unpurified use at 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
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WB | (7) |
1/10000. Predicted molecular weight: 14 kDa.
For unpurified use at 1/1000.00000 - 1/10000.00000 |
IHC-P |
1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
For unpurified use at 1/15 to 1/300. |
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IP |
1/600.
For unpurified use at 1/50 |
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ICC/IF | (1) |
Use a concentration of 0.1 µg/ml.
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Notes |
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Flow Cyt (Intra)
1/200. For unpurified use at 1/20. ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody. |
WB
1/10000. Predicted molecular weight: 14 kDa. For unpurified use at 1/1000.00000 - 1/10000.00000 |
IHC-P
1/150. Perform heat mediated antigen retrieval before commencing with IHC staining protocol. For unpurified use at 1/15 to 1/300. |
IP
1/600. For unpurified use at 1/50 |
ICC/IF
Use a concentration of 0.1 µg/ml. |
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
- Omim: 163890 Human
- SwissProt: P37840 Human
- Unigene: 21374 Human
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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 [MJFR1] (ab138501) at 1/10000 dilution
Lane 1 : Wild-type HEK-293T cell lysate
Lane 2 : SNCA knockout HEK-293T cell lysate
Lysates/proteins at 20 µg per lane.
Performed under reducing conditions.
Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Lanes 1- 2: Merged signal (red and green). Green - ab138501 observed at 18 kDa. Red - Anti-GAPDH antibody [6C5] - Loading Control (ab8245) observed at 37 kDa.
ab138501 was shown to react with SNCA in wild-type HEK-293T cells in western blot. Loss of signal was observed when knockout cell line ab255433 (knockout cell lysate ab263769) was used. Wild-type HEK-293T and SNCA knockout HEK-293T cell lysates were subjected to SDS-PAGE. Membrane was blocked for 1 hour at room temperature in 0.1% TBST with 3% non-fat dried milk. ab138501 and Anti-GAPDH antibody [6C5] - Loading Control (ab8245) overnight at 4°C at a 1 in 10000 dilution and a 1 in 20000 dilution respectively. Blots were developed with Goat anti-Rabbit IgG H&L (IRDye®800CW) preadsorbed (ab216773) and Goat anti-Mouse IgG H&L (IRDye®680RD) preadsorbed (ab216776) secondary antibodies at 1 in 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 [MJFR1] (ab138501)
IHC image of alpha Synuclein staining in normal Human cerebral cortex formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab138501, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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ab138501 staining Alpha-Synuclein in wild-type Hap1 cells (top panel) and SNCA knockout Hap1 cells (bottom panel). The cells were fixed with 4% paraformaldehyde (10 min) then permeabilized with 0.1% Triton X-100 for 5 minutes and then blocked with 1% BSA/10% normal goat serum/0.3M glycine in 0.1% PBS-Tween for 1h. The cells were then incubated with ab138501 at 0.1 μg/ml concentration and ab7291 (Mouse monoclonal to alpha Tubulin) at 1/1000 dilution overnight at 4°C followed by a further incubation at room temperature for 1h with a goat secondary antibody to rabbit IgG (Alexa Fluor® 488) (ab150081) at 2 μg/ml (shown in green) and a goat secondary antibody to mouse IgG (Alexa Fluor® 594) (ab150120) at 2 μg/ml (shown in red). Nuclear DNA was labelled in blue with DAPI.
Image was taken with a confocal microscope (Leica-Microsystems TCS SP8). -
Immunocytochemistry/Immunofluorescence analysis of U87-MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling alpha Synuclein with purified ab138501 at 1/150 dilution (left panel). Cells were fixed with 4% paraformaldehyde. A Goat anti rabbit IgG(Alexa Fluor®555) (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain (right panel).
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All lanes : Anti-Alpha-synuclein antibody [MJFR1] (ab138501) at 1/10000 dilution
Lane 1 : Wild-type HAP1 whole cell lysate
Lane 2 : SNCA knockout HAP1 whole cell lysate
Lane 3 : Human brain whole cell lysate
Lane 4 : Mouse brain whole cell lysate
Lysates/proteins at 40 µg per lane.
Predicted band size: 14 kDaLanes 1 - 4: Merged signal (red and green). Green - ab138501 observed at 14 kDa. Red - loading control, ab8245, observed at 37 kDa.
ab138501 was shown to specifically react with SNCA in wild-type HAP1 cells. No band was observed when SNCA knockout samples were used. Wild-type and SNCA knockout samples were subjected to SDS-PAGE. Ab138501 and ab8245 (Mouse anti GAPDH loading control) were incubated overnight at 4°C at 1/10000 and 1/10000 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 1/10000 dilution for 1 hour at room temperature before imaging.
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Overlay histogram showing HAP1 wildtype (green line) and HAP1-SNCA knockout cells (red line) stained with ab138501. The cells were fixed with 80% methanol (5 min) and then permeabilized with 0.1% PBS-Triton X-100 for 15 min. The cells were then incubated in 1x PBS / 10% normal goat serum to block non-specific protein-protein interactions followed by the antibody (ab138501, 1µg/ml) for 30 min at 22°C. The secondary antibody used was Alexa Fluor® 488 goat anti-rabbit IgG (H&L) presorbed (ab150081) at 1/2000 dilution for 30 min at 22°C. A rabbit IgG isotype control antibody (ab172730) was used at the same concentration and conditions as the primary antibody (HAP1 wildtype - black line, HAP1-SNCA knockout - grey line). Unlabelled sample was also used as a control (this line is not shown for the purpose of simplicity). Acquisition of >5,000 events were collected using a 50 mW Blue laser (488nm) and 530/30 bandpass filter.
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Anti-Alpha-synuclein antibody [MJFR1] (ab138501) at 1/10000 dilution (purified) + Human fetal brain at 20 µg
Secondary
Anti-Rabbit IgG (HRP), specific to the non-reduced form of IgG at 1/1000 dilution
Predicted band size: 14 kDa
Observed band size: 18 kDa why is the actual band size different from the predicted?Blocking and diluting buffer and concentration: 5% NFDM/TBST
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Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling alpha Synuclein with purified ab138501 at 1/200 dilution. The secondary antibody was Goat anti rabbit IgG (FITC) at 1/150 dilution.
The Isotype control is Rabbit monoclonal IgG (green line).
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Immunocytochemistry/Immunofluorescence analysis of U87-MG (Human glioblastoma-astrocytoma epithelial cell line) cells labeling alpha Synuclein with unpurified ab138501 at 1/15 dilution (left panel). Cells were fixed with 4% paraformaldehyde. A Goat anti rabbit IgG(Alexa Fluor®555) (1/200) was used as the secondary antibody. DAPI (blue) was used as the nuclear counter stain (right panel).
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
IHC image of alpha Synuclein staining in Normal human Substantia Nigra formalin fixed paraffin embedded tissue section, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab138501, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
IHC image of alpha Synuclein staining in Parkinson Human Substantia Nigra formalin fixed paraffin embedded tissue section*, performed on a Leica Bond™ system using the standard protocol F. The section was pre-treated using heat mediated antigen retrieval with sodium citrate buffer (pH6, epitope retrieval solution 1) for 20 mins. The section was then incubated with ab138501, 5µg/ml, for 15 mins at room temperature and detected using an HRP conjugated compact polymer system. DAB was used as the chromogen. The section was then counterstained with haematoxylin and mounted with DPX.
For other IHC staining systems (automated and non-automated) customers should optimize variable parameters such as antigen retrieval conditions, primary antibody concentration and antibody incubation times.
*Tissue obtained from the Human Research Tissue Bank, supported by the NIHR Cambridge Biomedical Research Centre
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alpha Synuclein was immunoprecipitated from Human fetal brain tissue using purified ab138501 at 1/600 dilution. Western blot was performed from the immunoprecipitate using purified ab138501. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution. Blocking and dilution buffer and concentration: 5% NFDM/TBST.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney labeling alpha Synuclein with purified ab138501 at 1/150 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.
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alpha Synuclein was immunoprecipitated from Human fetal brain tissue using unpurified ab138501 at 1/50 dilution. Western blot was performed from the immunoprecipitate using unpurified ab138501. Goat Anti-Rabbit IgG, (H+L), Peroxidase conjugated was used as secondary antibody at 1/1000 dilution. Blocking and dilution buffer and concentration: 5% NFDM/TBST
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human clear cell carcinoma of kidney labeling alpha Synuclein with unpurified ab138501 at 1/15 dilution. Heat mediated antigen retrieval was performed using Tris/EDTA buffer pH 9. Prediluted HRP Polymer for Rabbit IgG was used as the secondary antibody. Counter stained with Hematoxylin.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human brain tissue labeling alpha Synuclein with unpurified ab138501 at 1/300 dilution.
Perform heat mediated antigen retrieval before commencing with IHC staining protocol.
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Intracellular Flow Cytometry analysis of 2% paraformaldehyde fixed HeLa (Human epithelial cells from cervix adenocarcinoma) cells labeling alpha Synuclein with unpurified ab138501 at 1/20 dilution. The secondary antibody was Goat anti rabbit IgG (FITC) at 1/150 dilution.
The Isotype control isRabbit monoclonal IgG (green line).
Protocols
Datasheets and documents
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SDS download
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Datasheet download
References (118)
ab138501 has been referenced in 118 publications.
- Laferrière F et al. Similar neuronal imprint and no cross-seeded fibrils in α-synuclein aggregates from MSA and Parkinson's disease. NPJ Parkinsons Dis 8:10 (2022). PubMed: 35027576
- Yu Z et al. Erythrocytic α-Synuclein Species for Parkinson's Disease Diagnosis and the Correlations With Clinical Characteristics. Front Aging Neurosci 14:827493 (2022). PubMed: 35185529
- Liu Z et al. α-Synuclein-containing erythrocytic extracellular vesicles: essential contributors to hyperactivation of monocytes in Parkinson's disease. J Neuroinflammation 19:53 (2022). PubMed: 35193594
- Liu W et al. Characterization of a Novel Monoclonal Antibody for Serine-129 Phosphorylated α-Synuclein: A Potential Application for Clinical and Basic Research. Front Neurol 13:821792 (2022). PubMed: 35250825
- Wu Z et al. Prognostic significance of SNCA and its methylation in bladder cancer. BMC Cancer 22:330 (2022). PubMed: 35346107