Product Overview and Biological Background

Alpha-synuclein (α-Syn) is a 140-amino acid presynaptic protein encoded by the SNCA gene and is predominantly expressed in neural tissue. It plays roles in synaptic vesicle trafficking, dopamine regulation, and neuronal plasticity. Misfolding and aberrant aggregation of alpha-synuclein into oligomers and fibrils are central pathological events in Parkinson's disease (PD), Lewy body dementia, and multiple system atrophy—collectively termed synucleinopathies.

Phosphorylation is among the most functionally important post-translational modifications (PTMs) of alpha-synuclein. While phosphorylation at Ser129 is extensively studied and found highly enriched in Lewy bodies, tyrosine phosphorylation—particularly at Tyr125—has emerged as a biologically relevant modification. Tyr125 is located in the C-terminal acidic tail of alpha-synuclein, a region that modulates aggregation propensity and protein–protein interactions. Phosphorylation at this residue has been reported to influence synuclein conformation, membrane binding, and aggregation kinetics.

Recombinant Human Alpha-Synuclein Protein (Phospho-Tyr125) (SKU: rp377730) from Aladdin Scientific provides researchers with a defined, site-specifically phosphorylated form of the full-length human protein for use in biochemical, biophysical, and cell-based assays. The manufacturer reference for this product is absea#PC-062.

Handling, Reconstitution, and Storage

Proper handling of Recombinant Human Alpha-Synuclein Protein (Phospho-Tyr125) is critical to preserving phosphorylation integrity and biological activity. Follow the guidelines below:

• Upon receipt: Inspect the vial for integrity. Briefly centrifuge before opening to collect lyophilized material (if supplied lyophilized) or ensure liquid formulation is fully thawed on ice.
• Reconstitution (lyophilized form): Reconstitute in sterile, ultra-pure water or the buffer specified in the product datasheet to the recommended concentration. Allow to dissolve gently; do not vortex vigorously, as mechanical shear can promote alpha-synuclein aggregation.
• Aliquoting: Prepare single-use aliquots immediately after reconstitution to minimize repeated freeze-thaw cycles, which can compromise phosphorylation status and promote aggregation.
• Storage: Store long-term at −80 °C. Short-term storage (up to a few days) at 4 °C may be acceptable depending on the formulation—consult the product-specific datasheet for confirmed conditions.
• Avoid: Repeated freeze-thaw cycles, elevated temperatures, strong agitation, and prolonged exposure to light.

Note: Specific buffer composition, protein concentration, and formulation details should be verified against the current product datasheet provided with the shipment (absea#PC-062).

Research Applications

Recombinant Human Alpha-Synuclein Protein (Phospho-Tyr125) is intended for research use only and is suitable for a range of in vitro applications:

• Biochemical aggregation assays: Investigate how Tyr125 phosphorylation modulates the kinetics of alpha-synuclein fibrillation compared to unmodified or Ser129-phosphorylated forms using Thioflavin T fluorescence assays or atomic force microscopy (AFM).
• Antibody validation and Western blotting: Use as a positive control antigen for validating phospho-Tyr125-specific antibodies in Western blot, dot blot, or ELISA formats.
• Protein–protein interaction studies: Examine how C-terminal Tyr125 phosphorylation affects interactions with binding partners such as synphilin-1, 14-3-3 proteins, or components of the ubiquitin-proteasome system.
• Structural and biophysical studies: Employ in NMR, circular dichroism (CD), or dynamic light scattering (DLS) experiments to characterize conformational changes induced by phosphorylation.
• Cell-based assays: Introduce exogenously into neuronal cell models to study uptake, seeding competency, and downstream signaling events relevant to PD pathology.
• Enzyme and kinase activity assays: Serve as a substrate reference or product standard for tyrosine kinase assays targeting alpha-synuclein.

Researchers should empirically optimize protein concentrations, buffer conditions, and assay formats for each specific experimental application.

Quality, Purity, and Regulatory Notes

Recombinant Human Alpha-Synuclein Protein (Phospho-Tyr125) (SKU: rp377730) is manufactured and quality-controlled to meet research-grade specifications. Key quality considerations include:

• Phosphorylation site specificity: The protein is specifically phosphorylated at Tyr125 of the human alpha-synuclein sequence. Confirm phosphorylation verification method (e.g., mass spectrometry) in the product datasheet.
• Purity: Research-grade recombinant proteins are typically assessed by SDS-PAGE and may include endotoxin testing. Refer to the Certificate of Analysis (CoA) provided with each lot for confirmed purity values.
• Intended use: This product is for in vitro research use only. It is not approved for diagnostic, therapeutic, or clinical use in humans or animals.
• Lot-to-lot variation: Researchers should consult the lot-specific CoA for precise molecular weight, concentration, purity percentage, and activity data.

For regulatory, safety, and disposal information, refer to the Safety Data Sheet (SDS) available from Aladdin Scientific upon request.