Anti-Alpha-synuclein antibody [LB 509] (ab27766)

Overview

  • Product name
    Anti-Alpha-synuclein antibody [LB 509]
    See all Alpha-synuclein primary antibodies
  • Description
    Mouse monoclonal [LB 509] to Alpha-synuclein
  • 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.

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

Anti-Alpha-synuclein antibody [LB 509] images

  • IHC-P image of Alpha Synuclein (ab27766) staining in human brain samples with Multiple Systems Atrophy. 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

  • 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

  • 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

  • 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
    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
  • 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

    Predicted band size : 14 kDa
  • 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.

References for Anti-Alpha-synuclein antibody [LB 509] (ab27766)

This product has been referenced in:

See all 14 Publications for this product

Product Wall

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
Username

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
Username

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
Username

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
Username

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
Username

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
Username

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%
Username

Abcam user community

Verified customer

Submitted Apr 16 2013

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

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.
<...

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Thanks for sending the updated blot.

I believe its expected that Synuclein should be detected in the rat brain extracts, which it is not. There may be a band in your overexpressed Synuclein protein but it is difficult to determine with the la...

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