Alexa Fluor™ 647-Labeled Human Alpha-Synuclein Pre-formed Fibrils Protein, His Tag

Synonym

SNCA, NACP, PARK1, alpha-Synuclein

Source

Alexa Fluor 647-Labeled Human Alpha-Synuclein Pre-formed Fibrils Protein, His Tag (ALN-HA1H4) is expressed from E. coli cells. It contains AA Met 1 - Ala 140 (Accession # P37840-1).

Predicted N-terminus: Met 1

Molecular Characterization

Other Tags and Version Biotin & Other Labeled Version

This protein carries a polyhistidine tag at the C-terminus.

The protein has a calculated MW of 16.3 kDa.

Conjugate

AF647

Excitation Wavelength: 640 nm

Emission Wavelength: 672 nm

Labeling

The primary amines in the side chains of lysine residues and the N-terminus of the protein are conjugated with AF647 using standard chemical labeling method. The residual AF647 is removed by molecular sieve treatment during purification process.

Protein Ratio

The AF647 to protein molar ratio is 0.2-0.4.

Application

1. Sonication Conditions, Dissolution, Aliquoting, Storage, and Notes for PFF

Thawing: Thaw PFFs rapidly in a 37 °C water bath, or allow to thaw at room temperature.

Aliquoting: Since PFFs are supplied as a suspension, pipette up and down thoroughly before aliquoting to ensure homogeneity.

Storage: Store at -80 °C at all times; avoid storage at 4 °C or -20 °C, which can induce fibril depolymerization. * α-Syn fibrils cold-denatured to monomers at 0-20 °C and heat-denatured at 60-110 °C.

Sonication: The sonication protocols below are based on cell-based assay conditions.

-Option 1 (Probe Sonicator): Use a probe sonicator (SCIENTZ) at 10% power (-95 W), applying 60 pulses of 0.5 s on/0.5 s off (recommended).

-Option 2 (Ultrasonic Bath): Sonicate in an ultrasonic cleaner (40 kHz, 200-400 W) at 37 °C for 1 hour; avoid performing the treatment at 4 °C or 20 °C.

Note: The above sonication protocols are based on cell-based assays; other applications (e.g., in vivo injections) may require optimization-users should determine their optimal settings for their specific use case.

2. Recommendations for PFF Use in Animal Models

Minimize Freeze-Thaw Cycles: For animal studies, it is recommend to use the PFF at once or aliquot before use to avoid repeated freeze-thaw cycles.

PFF Maintenance During Injections: During stereotaxic injections, keep the sonicated PFFs in a 37 °C water bath to prevent re-aggregation or sedimentation.

Purity

>90% as determined by SDS-PAGE.

Formulation

Supplied as 0.2 μm filtered solution in PBS, pH7.4 with trehalose as protectant.

Contact us for customized product form or formulation.

Shipping

This product is supplied and shipped with dry ice, please inquire the shipping cost.

Storage

Please protect from light and avoid repeated freeze-thaw cycles.

This product is stable after storage at:

1. The product should be stored at -70°C or room temperature for short storage. Do not store fibrils on ice or at 4°C;
2. The unsonicated fibril is validated to be stable after storage at -70°C for 1 year under sterile conditions;
3. The sonicated fibril should be stored at -70°C for not more than 8 weeks.

ACRO Quality Management System

1. QMS(ISO, GMP)
2. Quality Advantages
3. Quality Control Process