Alpha-Synuclein Fibrils, Human

Cosmo Bio

Catalog No.: CSR-SYN03

Product Description

This product is sonicated, preformed human α-synuclein fibrils confirmed to function as seed fibrils in the α-Synuclein Aggregation Assay Kit (CSR-SYN01).

Source: Human
Expression System: E.coli
Quantity: 0.1mg
Concentration: 1mg/mL
Purity: ≥ 90% (SDS-Page)
Storage: -80°C

Related Products

Product	Cat. No.
Alpha-Synuclein Aggregation Assay Kit	CSR-SYN01
Amyloid Fluorescent Staining Kit	CSR-SYN02
Alpha-Synuclein Fibrils, Human	CSR-SYN03
Alpha-Synuclein, Human	CSR-SYN04
Alpha-Synuclein Fibrils, Mouse	CSR-SYN05
Alpha-Synuclein, Recombinant, Mouse	CSR-SYN06

Citations for Alpha-Synuclein Fibrils, Human – 3 Found

Inoue, Eiji; Suzuki, Takahiro; Shimizu, Yasuharu; Sudo, Keiichi; Kawasaki, Haruhisa; Ishida, Norio. Saffron ameliorated motor symptoms, short life span and retinal degeneration in Parkinson's disease fly models. Gene. 2021;799:145811. 

Karaki, Tatsuya; Haniu, Hisao; Matsuda, Yoshikazu; Tsukahara, Tamotsu. Lysophospholipids-potent candidates for brain food, protects neuronal cells against α-Synuclein aggregation. Biomedicine & Pharmacotherapy = Biomedecine & Pharmacotherapie. 2022;156:113891. 

Inoue, Eiji; Suzuki, Takahiro; Shimizu, Yasuharu; Sudo, Keiichi; Kawasaki, Haruhisa; Ishida, Norio. Saffron ameliorated motor symptoms, short life span and retinal degeneration in Parkinson's disease fly models. Gene. 2021;799:145811. 

Alpha-Synuclein Fibrils, Human FAQs

Does Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) contain monomers?
Following fibrillation, almost no monomers were detected in the supernatant. Furthermore, two rounds of ultracentrifugation are performed during manufacturing to remove any remaining non-fibrillized protein. Therefore, almost no monomers are expected to remain in the final product.

What is the molecular weight of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03)?
The molecular weight of Human Alpha-Synuclein Fibrils cannot be determined because this product consists of multimers and polymers.

What are the amino acid sequences of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) and Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) ?
Both products have the same amino acid sequence (140 amino acids):
MDVFMKGLSKAKEGVVAAAEKTKQGVAEAAGKTKEGVLYVGSKTKEGVVHGVATVAEKTKEQVTNVGGAV
VTGVTAVAQKTVEGAGSIAAATGFVKKDQLGKNEEGAPQEGILEDMPVDPDNEAYEMPSEEGYQDYEPEA

Can I purchase large quantities of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) from a single lot?
Yes. We can supply up to 40 vials (4 mg total) of Human Alpha-Synuclein Fibrils from a single lot. Please contact us for pricing and availability information regarding bulk orders.

Can I detect Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) and Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) using a phospho-specific antibody?
No. Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) is not phosphorylated, and Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) are prepared by fibrillization of this non-phosphorylated protein. Therefore, neither product is expected to be detected by phospho-specific α-synuclein antibodies. If a signal is observed in Western blot analysis, it is likely due to non-specific cross-reactivity of the antibody.

When Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) is dissolved in SDS sample buffer and analyzed by Western blotting, a band corresponding to the monomer molecular weight is observed. Does this indicate the presence of monomers?
When Human Alpha-Synuclein Fibrils is dissolved in SDS sample buffer and analyzed by Western blotting, a band at the monomer molecular weight (14.5 kDa) may appear. This does not necessarily indicate the presence of monomers in the sample. It is unlikely that monomers remain after purification or are generated during sonication. The observed band most likely results from a small amount of monomer released during SDS treatment.

How is Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) prepared?
Human Alpha-Synuclein Fibrils is prepared as follows:
1. Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) is fibrillized at 37°C.
2. The sample is ultracentrifuged, and the fibrils are collected as a pellet. Non-fibrillized monomers remain in the supernatant and are removed.
3. Physiological saline is added to the pellet, followed by a second ultracentrifugation step to further remove residual monomers.
4. The fibrils are resuspended in physiological saline and subjected to sonication. The resulting preparation is designated as Human Alpha-Synuclein Fibrils.

How should I dispose of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03), and why are specific autoclaving conditions required?
When disposing of Human Alpha-Synuclein Fibrils, please inactivate the fibrils using one of the following methods as specified in the datasheet:
Autoclave at 134°C for 20 minutes, OR
Autoclave at 121°C for 20 minutes in the presence of ≥ 3% SDS.

Why standard autoclaving (121°C, 20 min) is insufficient:
Standard autoclaving (121°C, 20 min) alone is not enough to completely eliminate the seeding activity of α-synuclein fibrils. The enhanced conditions (134°C or addition of SDS) are necessary to ensure full inactivation.
For data showing that seed activity (aggregation ability) is lost by autoclaving at 134°C for 20 minutes, or by adding SDS (3% or more) followed by autoclaving at 121°C for 20 minutes.
Please refer to the following publication.

Reference:
Degradation and inactivation of α-synuclein strains
Luc Bousset et al. Acta Neuropathol Commun. 2018 Apr 18;6(1):31. doi: 10.1186/s40478-018-0535-z.

How is the seeding activity of Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03) evaluated?
Seeding activity is evaluated by adding 1 µg of Human Alpha-Synuclein Fibrils to 100 µL of Recombinant Human Alpha-Synuclein (E. coli expressed) (Cat. No. CSR-SYN04) and monitoring Thioflavin T fluorescence over time. Seeding activity is confirmed when the fluorescence intensity exceeds the threshold after 15–20 hours of incubation.

Do you have endotoxin level data for Human Alpha-Synuclein Fibrils (Cat. No. CSR-SYN03)?
Measurements of endotoxin levels in Human Alpha-Synuclein Fibrils were conducted by a collaborating research group at the Tokyo Metropolitan Institute of Medical Science, which developed the production technology.
Please refer to the following publication.

Reference:
Exposure to bacterial endotoxin generates a distinct strain of α-synuclein fibril
Changyoun Kim et al. Sci Rep. 2016 Aug 4:6:30891. doi: 10.1038/srep30891.

For example, the endotoxin level was reported to be 10.6–11.3 endotoxin units per milligram of protein.