Human Alpha-Synuclein Pre-formed Fibrils Protein, Tag Free

SNCA, NACP, PARK1, alpha-Synuclein

Human Alpha-Synuclein Pre-formed Fibrils, Tag Free (ALN-H5115) is expressed from E. coli cells. It contains AA Met 1 - Ala 140 (Accession # P37840-1).

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.

Less than 1.0 EU per μg by the LAL method / rFC method.

>90% as determined by SDS-PAGE.

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

Contact us for customized product form or formulation.

This product is supplied and shipped on dry ice.

Please avoid repeated freeze-thaw cycles.

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