Anti-alpha Synuclein抗体[LB 509] (ab27766)

概述

  • 产品名称Anti-alpha Synuclein抗体[LB 509]
    参阅全部 alpha Synuclein 一抗
  • 描述
    小鼠单克隆抗体[LB 509] to alpha Synuclein
  • 特异性ab27766 recognises alpha synuclein. Recent western blot re-tests performed by our lab showed negative results in both mouse and rat. Therefore, we won't recommend this antibody in these species.
  • 经测试应用适用于: IHC-FoFr, ELISA, WB, IHC-Fr, IHC-P, Flow Cyt, ICCmore details
  • 种属反应性
    与反应: Human
  • 免疫原

    Lewy bodies purified from patients suffering dementia with Lewy bodies

  • 表位Ab27766 reacts with an epitope located in the region encoded by amino acids 115-122 of alpha synuclein.
  • 阳性对照
    • WB: full-length, recombinant alpha-synuclein protein.

性能

应用

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.

应用 Ab评论 说明
IHC-FoFr 1/500.
ELISA Use at an assay dependent concentration.
WB 1/100 - 1/10000. 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 Use at an assay dependent concentration.

靶标

  • 功能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.
  • 组织特异性Expressed principally in brain but is also expressed in low concentrations in all tissues examined except in liver. Concentrated in presynaptic nerve terminals.
  • 疾病相关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
  • 序列相似性Belongs to the synuclein family.
  • 结构域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.
  • 翻译后修饰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.
  • 细胞定位Cytoplasm, cytosol. Membrane. Nucleus. Cell junction, synapse. Secreted. Membrane-bound in dopaminergic neurons.
  • Information by UniProt
  • 数据库链接
  • 别名
    • 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
    • PD 1 antibody
    • PD1 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] 图像


  • Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 14 kDa


    Exposure time : 8 minutes
  • 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

  • ab27766 staining alpha Synuclein in Rat astrocytes by ICC/IF (Immunocytochemistry/immunofluorescence). Cells were fixed with paraformaldehyde, permeabilized with methanol and blocked with 5% BSA for 1 hour at 37°C. Samples were incubated with primary antibody (1/100) for 18 hours at 4°C. An Alexa Fluor® 488-conjugated Goat anti-mouse IgG polyclonal (1/200) was used as the secondary antibody.

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  • Overlay histogram showing PC12 cells stained with ab27766 (red line). The cells were fixed with methanol (5 min) and then permeabilized with 0.1% PBS-Tween for 20 min. The cells were then incubated in 1x PBS / 10% normal goat serum / 0.3M glycine to block non-specific protein-protein interactions followed by the antibody (ab27766, 1µg/1x106 cells) for 30 min at 22°C. The secondary antibody used was DyLight® 488 goat anti-mouse IgG (H+L) (ab96879) at 1/500 dilution for 30 min at 22°C. Isotype control antibody (black line) was mouse IgG1 [ICIGG1] (ab91353, 2µg/1x106 cells) used under the same conditions. Acquisition of >5,000 events was performed. This antibody gave a decreased signal in PC12 cells fixed with 4% paraformaldehyde (10 min)/permeabilized with 0.1% PBS-Tween used under the same conditions.

  • IHC-FoFr image of Alpha Synuclein staining on Synculein Transgenic mouse brain section stained using ab27766. The sections were incubated in 10% normal donkey serum in 0.1% PBS- and triton 0.3X100 for 1h to permeabilise the cells and block non-specific protein-protein interactions. The sections were then incubated with the antibody ab27766(1:500) overnight at +4°C. Mouse secondary was Alexa 488 and Granular ferritin expression (Alex 568) seen in the cytosol.

    See Abreview

  • Anti-alpha Synuclein antibody [LB 509] (ab27766) at 1/10000 dilution (Sample: ab51189,0.1ug)

    Secondary
    Goat Anti-Mouse IgG H&L (HRP) preadsorbed (ab97040) at 1/5000 dilution
    Developed using the ECL technique

    Performed under reducing conditions.

    Predicted band size : 14 kDa

Anti-alpha Synuclein antibody [LB 509] (ab27766)参考文献

This product has been referenced in:
  • Chai YJ  et al. Munc18-1 is a molecular chaperone for a-synuclein, controlling its self-replicating aggregation. J Cell Biol 214:705-18 (2016). Read more (PubMed: 27597756) »
  • Flores-Cuadrado A  et al. a-Synuclein staging in the amygdala of a Parkinson's disease model: cell types involved. Eur J Neurosci 41:137-46 (2015). IHC ; Mouse . Read more (PubMed: 25345880) »

See all 11 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

提交于 Feb 09 2015

Application Immunocytochemistry/ Immunofluorescence
Blocking step Serum as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 10% · Temperature: RT°C
Sample Human Cell (SH-SY5Y)
Specification SH-SY5Y
Permeabilization Yes - 0.5% Tween
Fixative Paraformaldehyde
Username

Abcam user community

Verified customer

提交于 Feb 09 2015

Abcam has not validated the combination of 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

提交于 May 13 2014

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Western blot
Loading amount 100 µg
Gel Running Conditions Reduced Denaturing
Sample Rat Tissue lysate - whole (brain tissue)
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

提交于 Sep 17 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

提交于 Sep 16 2013

Abcam guarantees this product to work in the species/application used in this Abreview.
Application Immunohistochemistry (PFA perfusion fixed frozen sections)
Blocking step BSA as blocking agent for 1 hour(s) and 0 minute(s) · Concentration: 5% · Temperature: 37°C
Sample Rat Tissue sections (Brain sections)
Specification Brain sections
Permeabilization Yes - Methanol
Fixative Acetone
Username

Abcam user community

Verified customer

提交于 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

提交于 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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