Recombinant Human Alpha-synuclein (mutated A53T) protein aggregate (Active) (ab256150)
Key features and details
- Expression system: Escherichia coli
- Purity: > 95% Ion Exchange Chromatography
- Active: Yes
- Suitable for: Functional Studies, IHC-P, Electron Microscopy
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Product name
Recombinant Human Alpha-synuclein (mutated A53T) protein aggregate (Active)
See all Alpha-synuclein proteins and peptides -
Biological activity
100 µM ab256149 seeded with 10 µM ab256150 in 25 µM Thioflavin T (PBS pH 7.4, 100 µL reaction volume) generated a fluorescence intensity of 28,000 Relative Fluorescence Units after incubation at 37°C with shaking at 600 rpm for 56 hours. Fluorescence was measured by excitation at 450 nm and emission at 485 nm on a Molecular Devices Gemini XPS microplate reader.
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Purity
> 95 % Ion Exchange Chromatography.
Certified >95% pure using SDS-PAGE analysis. -
Expression system
Escherichia coli -
Accession
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Protein length
Full length protein -
Animal free
No -
Nature
Recombinant -
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Species
Human -
Sequence
MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVH GVTTVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQL GKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA -
Predicted molecular weight
14 kDa -
Amino acids
1 to 140 -
Modifications
mutated A53T -
Additional sequence information
NP_000336.1
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Description
Recombinant human Alpha-synuclein (mutated A53T) protein (Active)
Preparation and Storage
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Alternative names
- Alpha synuclein
- Alpha-synuclein
- Alpha-synuclein, isoform NACP140
see all -
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
modificationsPhosphorylated, 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
Images
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Functional Studies - Recombinant Human Alpha-synuclein (mutated A53T) protein aggregate (Active) (ab256150)
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 a limited increase in fluorescence (correlated to alpha synuclein aggregation) over time in A53T alpha synuclein monomers (ab256149). A much greater increase in fluorescence is seen when 100 µM monomer (ab256149) is combined with 10 µM of fibrils (ab256150) as the fibrils seed the formation of new fibrils from the pool of active monomers. Thioflavin T ex = 450 nm, em = 485 nm.
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Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Recombinant Human Alpha-synuclein (mutated A53T) protein aggregate (Active) (ab256150)Immunohistochemical analysis of primary rat hippocampal neurons showing lewy body inclusion formation when treated with A53T mutant Alpha Synuclein Protein Preformed Fibrils (ab256150) (B) but not when treated with a media control (A). Tissue: Primary hippocampal neurons. Species: Sprague-Dawley rat. Primary Antibody: Rabbit anti-pSer129 Antibody. Fibrils were diluted to 1 ug/uL in neuronal media consisting of B27, Glutamax, penicillin/strip, and neurobasalA and sonicated for 1 hour in a water bath. The sonicated stock was then used to achieve the final concentration of 1 ug/mL in the wells.
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Electron Microscopy - Recombinant Human Alpha-synuclein (mutated A53T) protein aggregate (Active) (ab256150)TEM of ab256150.
