The application notes include recommended starting dilutions; optimal dilutions/concentrations should be determined by the end user.
Endogenous alpha-synuclein phosphorylation. 100 µM alpha synuclein protein monomer (ab218818) seeded with 10 nM 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.
>95% by SDS-PAGE . ab218819 was purified by ion-exchange.
Snca synuclein, alpha (non A4 component of amyloid precursor)
Synuclein alpha 140
Synuclein, alpha (non A4 component of amyloid precursor)
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.
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
Belongs to the synuclein family.
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.
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.
Functional Studies - Recombinant human Alpha-synuclein protein aggregate (Active) (ab218819)
ThT emission curves show increased fluorescence (correlated to alpha-synuclein protein aggregation) over time when 10 nM 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 nM of control alpha-synuclein aggregate (purple line), or 100 µM of control alpha-synuclein monomer (ab218816) is combined with 10 nM of control alpha-synuclein aggregate (ab218817) (dark blue). ThT ex = 450 nm, em = 485 nm. View protocol.
Immunohistochemistry (Formalin/PFA-fixed paraffin-embedded sections) - Recombinant human Alpha-synuclein protein aggregate (Active) (ab218819)
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.
SDS-PAGE - Recombinant human Alpha-synuclein protein aggregate (Active) (ab218819)
SDS-PAGE analysis of ab218819.
has not yet been referenced specifically in any publications.
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