Overview

  • 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: WB, IHC-P, Flow Cyt, IP, ELISA, ICC/IFmore details
  • Species reactivity
    Reacts with: Human
  • Immunogen

    Recombinant full length protein corresponding to Human Alpha-synuclein aa 1 to the C-terminus.
    Database link: P37840

  • Epitope
    The epitope was mapped to amino acids 118-123 (VDPDNE).
  • Positive control
    • WB: Recombinant Human Alpha-synuclein protein (ab51189), HAP1 (WT), HAP1 (SNCA KO) whole 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: Hap1-wt cells.
  • 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).

    Rat: We have preliminary internal testing data to indicate this antibody may not react with these species. Please contact us for more information.

    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.

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

    We are constantly working hard to ensure we provide our customers with best in class antibodies. As a result of this work we are pleased to now offer this antibody in purified format. We are in the process of updating our datasheets. The purified format is designated 'PUR' on our product labels. If you have any questions regarding this update, please contact our Scientific Support team.

    This product is a recombinant rabbit monoclonal antibody.

Properties

Applications

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
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.

See IHC antigen retrieval protocols.

Flow Cyt 1/200.

For unpurified use at 1/20.

ab172730 - Rabbit monoclonal IgG, is suitable for use as an isotype control with this antibody.

IP 1/600.

For unpurified use at 1/50

ELISA Use a concentration of 1 - 10 µg/ml.
ICC/IF 1/150.

For unpurified use at 1/15

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
    • 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, 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

Images

  • Lane 1: Wild type HAP1 whole cell lysate (40 µg)
    Lane 2: SNCA knockout HAP1 whole cell lysate (40 µg)
    Lane 3: Human brain whole cell lysate (40 µg)
    Lane 4: Mouse brain whole cell lysate (40 µg)

    Lanes 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.

  • 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.

  • 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).

  • 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.

  • 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).

  • 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

  • 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.

  • 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).

     

  • Anti-Alpha-synuclein antibody [MJFR1] (ab138501) at 1/1000 dilution (unpurified) + 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  

  • 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

  • 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.

  • 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.

  • 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

  • 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.

  • Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) analysis of Human brain tissue labeling alpha Synuclein with unpurified ab138501 at 1/300 dilution.

  • 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 is Rabbit monoclonal IgG (green line).

References

This product has been referenced in:
  • Abbineni PS  et al. Identification of ß-synuclein on secretory granules in chromaffin cells and the effects of a- and ß-synuclein on post-fusion BDNF discharge and fusion pore expansion. Neurosci Lett 699:134-139 (2019). Read more (PubMed: 30711526) »
  • Kruse N & Mollenhauer B Quantification of Alpha-Synuclein in Biological Fluids by Electrochemiluminescence-Based Detection. Methods Mol Biol 1948:59-68 (2019). Read more (PubMed: 30771170) »
See all 34 Publications for this product

Customer reviews and Q&As

1-5 of 5 Abreviews or Q&A

Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (synuclein inclusion in human frozen brain section)
Permeabilization
Yes - 0.2% triton in PBS
Specification
synuclein inclusion in human frozen brain section
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde + methanol

Diego Perez Rodriguez

Verified customer

Submitted Dec 03 2018

Application
Western blot
Sample
Human Recombinant protein (Human and transgenic mouse M83)
Gel Running Conditions
Non-reduced Non-Denaturing (Native)
Loading amount
1 µg
Specification
Human and transgenic mouse M83
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 22°C

Sophie Morgan

Verified customer

Submitted Jun 29 2018

Application
Western blot
Loading amount
30 µg
Gel Running Conditions
Reduced Denaturing (10% Gel)
Sample
Human Cell lysate - whole cell (SH-SY5Y Neuroblastoma cell line)
Specification
SH-SY5Y Neuroblastoma cell line
Blocking step
Milk as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 25°C

Mr. Fredrik Hoel

Verified customer

Submitted Dec 16 2014

Answer



ab138501 is an unpurified tissue culture supernatant product, so the concentration of the specific anti-alpha Synuclein antibody is not determined. However in general, for products of this type, the specific antibody concentration is about 0.05mg/ml.

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Answer


I have conducted a sequence comparison between the human and rat alpha synuclein proteins and there is a very high sequence homology. Out of the 140 amino acids of this protein, 133 amino acids are shared between human and rat which suggests that it is unlikely that any antibody you will find to alpha-synuclein will definitely not cross-react with rat since there is no sequence within this protein that varies significantly between rat and human.

ab138501 has been tested in mouse, rat and human and only showed positive results in mouse and human. This does not mean that it will not work in rat since with some optimisation, the rat protein could also be detected.

However, since our datasheet specifically states that this antibody does not work in rat, we will honour our guarantee and if you buy and test this antibody in rat and find that it works, we will refund or replace the antibody.

Read More

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