Overview

  • Product name
    Anti-Alpha-synuclein antibody [LB 509]
    See all Alpha-synuclein primary antibodies
  • Description
    Mouse monoclonal [LB 509] to Alpha-synuclein
  • Host species
    Mouse
  • Specificity
    This antibody recognizes human alpha-synuclein. Clone LB 509 recognizes amino acids 115-122 of alpha-synuclein and has been reported to be specific to human alpha-synuclein (Jakes et al., Neurosci Lett., 1999). Therefore, we only guarantee this antibody to detect human alpha-synuclein. However, several customer Abreviews have demonstrated positive staining in rodent samples.
  • Tested applications
    Suitable for: IHC-FoFr, ELISA, WB, IHC-Fr, IHC-P, Flow Cyt, ICC/IF, ICCmore details
  • Species reactivity
    Reacts with: Rat, Human
  • Immunogen

    Lewy bodies purified from patients suffering dementia with Lewy bodies

  • Epitope
    Ab27766 reacts with an epitope located in the region encoded by amino acids 115-122 of alpha-synuclein.
  • Positive control
    • WB: full-length, recombinant alpha-synuclein protein.
  • General notes

    This antibody clone is manufactured by Abcam.

    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.

    If you require this antibody in a particular buffer formulation or a particular conjugate for your experiments, please contact orders@abcam.com or you can find further information here.

Properties

Applications

Our Abpromise guarantee covers the use of ab27766 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-FoFr 1/500.
ELISA Use at an assay dependent concentration.
WB 1/100 - 1/1000. Predicted molecular weight: 14 kDa.
IHC-Fr 1/100 - 1/1000.
IHC-P 1/100 - 1/1000. Do not perform antigen retrieval.
Flow Cyt Use 1µg for 106 cells.

ab170190 - Mouse monoclonal IgG1, is suitable for use as an isotype control with this antibody.

ICC/IF Use at an assay dependent concentration.
ICC Use at an assay dependent concentration.

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

  • All lanes : Anti-Alpha-synuclein antibody [LB 509] (ab27766) at 5 µg/ml

    Lane 1 : Recombinant Human Alpha-synuclein protein (ab51189)
    Lane 2 : Human brain hippocampus tissue lysate - total protein (ab30180)

    Lysates/proteins at 10 µg per lane.

    Secondary
    All lanes : Goat polyclonal to Mouse IgG - H&L - Pre-Adsorbed (HRP) at 1/5000 dilution

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 14 kDa
    Observed band size: 16 kDa
    why is the actual band size different from the predicted?
    Additional bands at: 32 kDa (possible dimer)


    Exposure time: 20 minutes
  • IHC-P image of Alpha Synuclein (ab27766) staining in human brain samples from patients with Multiple Systems Atrophy (MSA). The sections were subjected to heat-mediated antigen retrieval with citrate buffer. In addition, some slides received a 15 minute pre-treatment with Formic Acid. Sections were incubated in 20% serum for 30 minutes at +18°C to block non-specific protein-protein interactions. The sections were then incubated with ab27766 (1:400) for one hour at +18°C, followed by Biotin conjugated anti-mouse goat secondary antibody (1/200). Formic acid pre-treatment (15min) revealed more inclusions in MSA tissue.

    See Abreview

  • Human neuroblastoma cells stained for alpha-synuclein (green) using ab27766 in immunofluorescence. The neuroblastoma cells were fixed with paraformaldehyde and incubated with ab27766 (used at 5 μg/ml) for 12 hours at 4°C. A FITC conjugated Goat anti-Mouse IgG secondary antibody was then used.

    See Abreview

  • Overlay histogram showing PC12 (NGF differentiated) cells stained with ab27766 (red line). The cells were fixed with 4% formaldehyde (10 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 (ab27766, 1µg/1x106) for 30 min at 22°C. The secondary antibody used was Goat Anti-Mouse IgG H&L (Alexa Fluor® 488) preadsorbed (ab150117) at 1/2000 dilution for 30 min at 22°C.

    Isotype control antibody (black line) was Mouse IgG1 [15-6E10A7] (ab170190) used at the same concentration and conditions as the primary antibody. Unlabelled sample (blue line) was also used as a control.

    Acquisition of >5,000 events were collected using a 50mW Blue laser (488nm) and 530/30 bandpass filter.

  • SH-SY5Y neuroblastoma cells stained for alpha-synuclein (green) using ab27766 in immunofluorescence. SH-SY5Y cells were fixed with paraformaldehyde, permeabilized with 0.5% Tween-20 and blocked with 10% serum for 1 hour at room temperature. Samples were incubated with ab27766 (diluted at 1/300) for 1 hour. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal was used as the secondary antibody (diluted at 1/200).

    See Abreview

  • Anti-Alpha-synuclein antibody [LB 509] (ab27766) at 1/1000 dilution + Recombinant Human Alpha-synuclein protein (ab51189) at 0.1 µg

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 5000 µg

    Developed using the ECL technique.

    Performed under reducing conditions.

    Predicted band size: 14 kDa


    Exposure time: 8 minutes

References

This product has been referenced in:
  • Gerson JE  et al. Tau oligomers mediate a-synuclein toxicity and can be targeted by immunotherapy. Mol Neurodegener 13:13 (2018). Read more (PubMed: 29544548) »
  • Finkelstein DI  et al. The novel compound PBT434 prevents iron mediated neurodegeneration and alpha-synuclein toxicity in multiple models of Parkinson's disease. Acta Neuropathol Commun 5:53 (2017). WB ; Human . Read more (PubMed: 28659169) »
See all 24 Publications for this product

Customer reviews and Q&As

1-10 of 18 Abreviews or Q&A

Application
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections)
Sample
Dog Tissue sections (canine brain)
Antigen retrieval step
Heat mediated - Buffer/Enzyme Used: citrate buffer
Permeabilization
Yes - 0.025% TritonX
Specification
canine brain
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: 25°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 23 2018

Application
Western blot
Loading amount
25 µg
Gel Running Conditions
Reduced Denaturing
Sample
Human Cell lysate - whole cell (SH-SY5Y)
Specification
SH-SY5Y
Treatment
50 ´M, 100 ´M or 200 ´M of dopamine with or without 100 nM of MG132 proteasome inhibitor for 72 h
Blocking step
Milk as blocking agent for 24 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: RT°C

Dr. B Roberts

Verified customer

Submitted Feb 09 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (SH-SY5Y)
Permeabilization
Yes - 0.5% Tween
Specification
SH-SY5Y
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Feb 09 2015

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Human Cell (neuroblastoma)
Permeabilization
No
Specification
neuroblastoma
Blocking step
Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 1% · Temperature: 24°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted May 13 2014

Abcam has not validated the combination of species/application used in this Abreview.
Application
Western blot
Sample
Rat Tissue lysate - whole (brain tissue)
Gel Running Conditions
Reduced Denaturing
Loading amount
100 µg
Specification
brain tissue
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C

Abcam user community

Verified customer

Submitted Sep 18 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunohistochemistry (PFA perfusion fixed frozen sections)
Sample
Rat Tissue sections (Brain sections)
Permeabilization
Yes - Methanol
Specification
Brain sections
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Acetone

Abcam user community

Verified customer

Submitted Sep 16 2013

Abcam has not validated the combination of species/application used in this Abreview.
Application
Immunocytochemistry/ Immunofluorescence
Sample
Rat Cell (Astrocytes)
Permeabilization
Yes - Methanol
Specification
Astrocytes
Blocking step
BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Fixative
Paraformaldehyde

Abcam user community

Verified customer

Submitted Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application
Western blot
Sample
Human Cell lysate - whole cell (Neuroblastoma)
Loading amount
15 µg
Specification
Neuroblastoma
Gel Running Conditions
Reduced Denaturing
Blocking step
(agent) for 1 hour(s) and 0 minute(s) · Concentration: 50%

Abcam user community

Verified customer

Submitted Apr 16 2013

Answer

Thank you for contacting us. The reviewwer used 90-100% formic acid for 10 minutes. Please let me kknow if you have further questions.

Read More

Answer

I am sorry this product did not perform as stated on the datasheet and for the inconvenience this has caused. As requested, I have issued a free of charge replacement for one vial of ab98929 and one vial of ab77028with the order number 1150942.

To check the status of the order please contact our Customer Service team and reference this number.

Please note that this free of charge replacement vial is also covered by our Abpromise guarantee. Should you still be experiencing difficulties, or if you have any further questions, please do not hesitate to let us know.

I wish you the best of luck with your research.

Read More

1-10 of 18 Abreviews or Q&A

Please note: All products are "FOR RESEARCH USE ONLY AND ARE NOT INTENDED FOR DIAGNOSTIC OR THERAPEUTIC USE"

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