重组人Alpha-synuclein蛋白aggregate (Active) (ab218819)
Key features and details
- Expression system: Escherichia coli
- Purity: > 95% SDS-PAGE
- Endotoxin level: <= 20.000 Eu/ml
- Active: Yes
- Suitable for: WB, Functional Studies, SDS-PAGE
描述
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产品名称
重组人Alpha-synuclein蛋白aggregate (Active)
参阅全部 Alpha-synuclein 蛋白酶 -
生物活性
Endogenous alpha-synuclein phosphorylation.
100 µM alpha synuclein protein monomer (ab218818) seeded with 10 µM alpha synuclein protein aggregate (ab218819) in 25 µM Thioflavin T (ab120751) (PBS pH 7.4, 100 µl reaction volume) generated a fluorescence intensity of 13,000 Relative Fluorescence Units after incubation at 37°C with shaking at 600 rpm for 24 hours.
Fluorescence was measured by excitation at 450 nm and emission at 485 nm on a microplate reader.
Endotoxin Level: 10-20 EU/mL
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纯度
> 95 % SDS-PAGE.
ab218819 was purified by ion-exchange. -
内毒素水平
<=20.000 Eu/ml -
表达系统
Escherichia coli -
Accession
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蛋白长度
Full length protein -
无动物成分
No -
性质
Recombinant -
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种属
Human -
序列
MDVFMKGLSK AKEGVVAAAE KTKQGVAEAA GKTKEGVLYV GSKTKEGVVH GVATVAEKTK EQVTNVGGAV VTGVTAVAQK TVEGAGSIAA ATGFVKKDQL GKNEEGAPQE GILEDMPVDP DNEAYEMPSE EGYQDYEPEA -
预测分子量
14 kDa -
氨基酸
1 to 140 -
额外的序列信息
(NP_000336.1) (GeneID 6622)
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描述
重组人Alpha-synuclein蛋白(Active)
相关产品
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Related Products
- Thioflavin T, Fluorescent cell-permeable amyloid binding benzothiazole salt (ab120751)
- Anti-Alpha-synuclein antibody [MJFR1] (ab138501)
- Anti-Alpha-synuclein antibody [4D6] - BSA and Azide free (ab1903)
- Recombinant Human Alpha-synuclein protein aggregate (Type 2) (ab218817)
- Recombinant Human Alpha-synuclein protein monomer (Active) (ab218818)
- Anti-Alpha-synuclein antibody [LB 509] (ab27766)
技术指标
Our Abpromise guarantee covers the use of ab218819 in the following tested applications.
The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
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应用
Western blot
Functional Studies
SDS-PAGE
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形式
Liquid -
补充说明
For best results, sonicate immediately prior to use. Sonication of PFFs is required in order to obtain fibrils with an average length of ~50nm, the optimal size for pathology.
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Concentration information loading...
制备和贮存
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稳定性和存储
Shipped on Dry Ice. Store at -80°C. Avoid freeze / thaw cycle.
Constituent: 95% PBS
This product is an active protein and may elicit a biological response in vivo, handle with caution.
常规信息
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别名
- Alpha synuclein
- Alpha-synuclein
- Alpha-synuclein, isoform NACP140
see all -
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
图片
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Immunohistochemical analysis of primary rat hippocampal neurons showing lewy body inclusion formation when treated with active Alpha Synuclein Protein Aggregate (ab218819) at 4 µg/ml (D-F), but not when treated with control Alpha Synuclein Protein Aggregate (ab218817) at 4 µg/ml (A-C). Tissue: Primary hippocampal neurons. Species: Sprague-Dawley rat. Fixation: 4% formaldehyde from PFA. Primary antibody: Mouse anti-pSer129 Antibody at 1/1000 24 hours at 4°C. Secondary antibody: FITC Goat Anti-Mouse (green) at 1/700 for 1 hour at RT. Counterstain: Hoechst (blue) nuclear stain at 1/4000 for 1 hour at RT. Localization: Lewy body incluscions. Magnification: 20x.
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ab218819 seeds the formation of new alpha synuclein fibrils from the pool of alpha synuclein monomers.
Thioflavin T is a fluorescent dye that binds to beta sheet-rich structures, such as those in alpha synuclein fibrils. Upon binding, the emission spectrum of the dye experiences a red-shift, and increased fluorescence intensity.
Thioflavin T emission curves show increased fluorescence (correlated to alpha synuclein protein aggregation) over time when 10 µM of ab218819 is combined with 100 µM of alpha synuclein monomer, as compared to ab218819 alone and alpha synuclein monomer alone.
Thioflavin T ex = 450 nm, em = 485 nm.
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TEM of active human alpha synuclein preformed fibrils (ab218819). Fibrils were sonicated and treated with uranyl acetate.
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SDS-PAGE analysis of ab218819.
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Immunohistochemistry analysis of rat brain injected with active human alpha synuclein PFFs (ab218819). Species: Female Sprague-Dawley Rat. Rat was injected with 2µL active human alpha synuclein PFFs (ab218819) in each of 2 injection sites: AP+1.6, ML+2.4, DV-4.2 from skull; and AP-1.4, ML+0.2, DV-2.8 from skull. 30-days post-injection. Fixation: Saline perfusion followed by 4% PFA fixation for 48 hrs. Secondary Antibody: Biotin-SP Donkey Anti-Rabbit IgG (H+L) at 1:500 for 2 hours in cold room with shaking. ABC signal amplification, DAB staining. Alpha synuclein pathology is seen in the striatum close to an injection site.
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TEM of ab218819.
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TEM of active human alpha synuclein preformed fibrils (ab218819) (top) and control (inactive) human alpha synuclein preformed fibrils (ab218817) (bottom). Fibrils were sonicated and treated with uranyl acetate. The active fibrils are shorter than the inactive fibrils.
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ThT emission curves show increased fluorescence (correlated to alpha-synucleinprotein aggregation) over time when 10 µM of active alpha-synuclein aggregate (ab218819) is combined with 100 µM of active alpha-synuclein monomer (ab218818) (light blue), as compared to when 100 µM of active alpha-synuclein monomer is combined with 10 µM of control alpha-synuclein aggregate (purple line), or 100 µM of control alpha-synuclein monomer (ab218816) is combined with 10 µM of control alpha-synuclein aggregate (ab218817) (dark blue). ThT ex = 450 nm, em = 485 nm. View protocol.
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TEM of active human alpha synuclein preformed fibrils (ab218819) (top) and control (inactive) human alpha synuclein preformed fibrils (ab218817) (bottom). Fibrils were sonicated and treated with uranyl acetate. The active fibrils are shorter than the inactive fibrils.
实验方案
To our knowledge, customised protocols are not required for this product. Please try the standard protocols listed below and let us know how you get on.
数据表及文件
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Datasheet download
文献 (6)
ab218819 被引用在 6 文献中.
- Uçar B et al. Spreading of Aggregated α-Synuclein in Sagittal Organotypic Mouse Brain Slices. Biomolecules 12:N/A (2022). PubMed: 35204664
- Ng L et al. Celastrol Downmodulates Alpha-Synuclein-Specific T Cell Responses by Mediating Antigen Trafficking in Dendritic Cells. Front Immunol 13:833515 (2022). PubMed: 35309340
- Tiwari S et al. UBA52 Is Crucial in HSP90 Ubiquitylation and Neurodegenerative Signaling during Early Phase of Parkinson's Disease. Cells 11:N/A (2022). PubMed: 36497031
- Piancone F et al. Inflammatory Responses to Monomeric and Aggregated a-Synuclein in Peripheral Blood of Parkinson Disease Patients. Front Neurosci 15:639646 (2021). PubMed: 33867921
- Baxter PS et al. Targeted de-repression of neuronal Nrf2 inhibits a-synuclein accumulation. Cell Death Dis 12:218 (2021). PubMed: 33637689
- Pediaditakis I et al. Modeling alpha-synuclein pathology in a human brain-chip to assess blood-brain barrier disruption. Nat Commun 12:5907 (2021). PubMed: 34625559