Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein

Novus Biologicals | Catalog # NBP2-54789

Novus Biologicals

Key Product Details

Source

E. coli

Applications

Immunohistochemistry, Western Blot, Immunocytochemistry/ Immunofluorescence, Bioactivity, Functional Assay, Microscopy, SDS-PAGE
View all alpha-Synuclein Proteins and Enzymes »

Product Specifications

Description

An un-tagged full length Human biologically active Alpha-Synuclein recombinant protein aggregate (pre-formed fibrils, Type 1), NCBI Accession #: NP_000336.1.

Source: E. coli

Amino Acid Sequence:

MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAVVTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA

Purity

Ion exchange chromatography

Predicted Molecular Mass

14.46 kDa.
Disclaimer note: The observed molecular weight of the protein may vary from the listed predicted molecular weight due to post translational modifications, post translation cleavages, relative charges, and other experimental factors.

Activity

Endogenous alpha-synuclein phosphorylation. 100 uM alpha synuclein protein monomer (NBP2-54788) seeded with 10 uM alpha synuclein protein PFF (NBP2-54789) in 25 uM Thioflavin T (PBS pH 7.4, 100 ul reaction volume) generated a fluorescence intensity of 13,000 Relative Fluorescence Units after incubation at 37 degrees C with shaking at 600 rpm for 24 hours. Fluorescence was measured by excitation at 450 nm and emission at 485 nm on a Molecular Devices Gemini XPS microplate reader.

Applications

Western Blot (1:100-1:2000)
SDS-Page (1:100-1:2000)

Protein / Peptide Type

Recombinant Protein

Scientific Data Images

Western Blot: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Western Blot: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Western Blot: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - Lane 1: Molecular Weight Ladder (MW). Lane 2: Active Alpha Synuclein Protein Aggregate.
Immunocytochemistry/ Immunofluorescence: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Immunocytochemistry/ Immunofluorescence: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Immunocytochemistry/Immunofluorescence: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - Primary rat hippocampal neurons show lewy body inclusion formation when treated with active Alpha Synuclein Protein Aggregate at 4 ug/ml (D-F), but not when treated with control Alpha Synuclein Protein Aggregate at 4 ug/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 4C. Secondary Antibody: FITC Goat Anti-Mouse (green) at 1:700 for 1 hours at RT. Counterstain: Hoechst (blue) nuclear stain at 1:4000 for 1 hour at RT. Localization: Lewy body incluscions. Magnification: 20x.
Immunohistochemistry: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Immunohistochemistry: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

Immunohistochemistry: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - Immunohistochemistry analysis of rat brain injected with Type 1 human alpha synuclein PFFs (NBP2-54789). Species: Female Sprague-Dawley Rat. Rat was injected with 16g Type 1 human alpha synuclein PFFs (NBP2-54789) 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. Primary antibody: rabbit monoclonal anti-pSer129 alpha synuclein. 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. Magnification: 20X. Alpha synuclein pathology is seen in the striatum close to an injection site.
In vitro assay: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

In vitro assay: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789]

In vitro assay: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - Thioflavin T emission curves show increased fluorescence (correlated to alpha Synuclein protein aggregation) over time when 10 nM of active alpha Synuclein aggregate (NBP2-54789) is combined with 100 uM of active alpha Synuclein monomer (NBP2-54788), as compared to active alpha Synuclein aggregate (NBP2-54789) and active alpha Synuclein monomer (NBP2-54788) alone. Thioflavin T ex = 450 nm, em = 485 nm.
Electron Microscopy: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - TEM of Type 1 alpha-Synuclein Pre-formed Fibrils (PFFs) (NBP2-54789)
Product Image: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - Toxicity results comparing Active Human Recombinant alpha-Synuclein Pre-formed Fibrils (Type 2) and Active Human Recombinant alpha-Synuclein Pre-formed Fibrils (Type 1) (Catalog No. NBP2-54789). Data was graphed after live cell imaging results were obtained using the following procedure: After 8 days in vitro, primary rat mixed cortical neuron cells were washed with 1X PBS and treated with 500 g/ml of Type 1 and Type 2 alpha-Synuclein Proteins for 20 hours at 37C. Following treatements, cells were washed with 2X PBS and incubated with a staining solution (2.0 M Cell Event + 2.5 M Ethidium homodimer + 2.5 g/ml Hoechst 33342 in sterile HBSS) for 30 minutes at 37C. The addition of the Type 2 alpha-Synuclein Proteins resulted in a significant increase in cell death.
Electron Microscopy: Recombinant Human alpha-Synuclein Active, Pre-formed Fibrils, (Type 1) Protein [NBP2-54789] - TEM of Type 1 alpha-Synuclein Pre-formed Fibrils (PFFs) (NBP2-54789)

Formulation, Preparation, and Storage

NBP2-54789
Formulation PBS
Concentration Please see the vial label for concentration. If unlisted please contact technical services.
Shipping The product is shipped with dry ice or equivalent. Upon receipt, store it immediately at the temperature recommended below.
Stability & Storage Store at -80C. Avoid freeze-thaw cycles.

Background: alpha-Synuclein

Alpha-synuclein, a member of the synuclein family, is a protein that was first identified in 1988 whose name is derived from its localization to both the synapse and nucleus (1-3). Specifically, it is expressed primarily in the brain, including Lewy Bodies (1-6). Alpha-synuclein is encoded by the SNCA gene, located on chromosome 4p21, and is processed as a 140 amino acid (aa) protein with a theoretical molecular weight of 14 kDa (1,2,4). Structurally alpha-synuclein consists of a N-terminal binding domain (1-60 aa), a central domain core region called the non-amyloid-beta component (NAC) (61-95 aa), and a C-terminal domain (96-140 aa) (1-3). The N-terminal region contains aa repeats with a KTKEGV consensus sequence that gives the protein its alpha-helical structure that associates with lipid membranes (1-4). The hydrophobic NAC region is responsible for alpha-synuclein aggregation and fibril formation (1-4). The acidic C-terminal tail is largely unstructured but can be targeted for post-translational modifications (1-4). The function of alpha-synuclein is not entirely understood, but it is shown to have a role in suppression of apoptosis, acting as a molecular chaperone, regulating glucose, and modulating calmodulin activity (1,3).

A number of studies have revealed that alpha-synuclein aggregation is a hallmark feature in a number of neurodegenerative diseases, referred to as synucleinopathies (2-4). Alpha-synuclein protein aggregates are a large component of Lewy bodies that are present in Parkinson's disease (PD), Lewy body dementia (LBD), and multiple system atrophy (1-6). Research has shown phosphorylation of alpha-synuclein at Ser129 moves the protein from the nucleus to the cytoplasm and promotes fibril formation associated with synucleinopathies (1,2,5). Recent studies also suggest that alpha-synuclein accumulation can prevent mitochondrial import machinery causing mitochondrial dysfunction that is often observed in neurodegeneration (5). It is thought that preventing alpha-synuclein aggregation may prevent PD, thus alpha-synuclein is a target for many potential therapeutic interventions aimed at decreasing aggregate formation or increasing clearance (1,2,4-6).

References

1. Villar-Pique, A., Lopes da Fonseca, T., & Outeiro, T. F. (2016). Structure, function and toxicity of alpha-synuclein: the Bermuda triangle in synucleinopathies. Journal of neurochemistry. https://doi.org/10.1111/jnc.13249

2. Emamzadeh F. N. (2016). Alpha-synuclein structure, functions, and interactions. Journal of research in medical sciences : the official journal of Isfahan University of Medical Sciences. https://doi.org/10.4103/1735-1995.181989

3. Burre J. (2015). The Synaptic Function of alpha-Synuclein. Journal of Parkinson's disease. https://doi.org/10.3233/JPD-150642

4. Lashuel, H. A., Overk, C. R., Oueslati, A., & Masliah, E. (2013). The many faces of alpha-synuclein: from structure and toxicity to therapeutic target. Nature reviews. Neuroscience. https://doi.org/10.1038/nrn3406

5. Rocha, E. M., De Miranda, B., & Sanders, L. H. (2018). Alpha-synuclein: Pathology, mitochondrial dysfunction and neuroinflammation in Parkinson's disease. Neurobiology of disease. https://doi.org/10.1016/j.nbd.2017.04.004

6. O'Leary, E. I., & Lee, J. C. (2019). Interplay between alpha-synuclein amyloid formation and membrane structure. Biochimica et biophysica acta. Proteins and proteomics. https://doi.org/10.1016/j.bbapap.2018.09.012

Alternate Names

NACP, PARK1, PARK4, SNCA, Synuclein-alpha

Gene Symbol

SNCA

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Product Documents

Certificate of Analysis

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Product Specific Notices

This product is for research use only and is not approved for use in humans or in clinical diagnosis. This product is guaranteed for 1 year from date of receipt.

Related Research Areas

Neuronal Lineage Markers

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Protocols

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FAQs

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  • Q: I'm looking for an alpha-Synuclein antibody with an epitope located in the first half (N-terminus) of the protein - preferably a monoclonal antibody. Can you help me with that?

    A: Please take a look at NB110-57475. It has been validated for human, rat and mouse and the applications ICC and WB and the epitope it detects is in the N terminal.

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