产品详情
分子别名(Synonym)
SNCA, NACP, PARK1, alpha-Synuclein
表达区间及表达系统(Source)
Human Alpha-Synuclein Pre-formed Fibrils, His Tag (ALN-H51H4) is expressed from E. coli cells. It contains AA Met 1 - Ala 140 (Accession # P37840-1).
Predicted N-terminus: Met 1
Request for sequence蛋白结构(Molecular Characterization)
This protein carries a polyhistidine tag at the C-terminus.
The protein has a calculated MW of 16.3 kDa.
应用说明(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.
内毒素(Endotoxin)
Less than 1.0 EU per μg by the LAL method / rFC method.
纯度(Purity)
>90% as determined by SDS-PAGE.
制剂(Formulation)
Supplied as 0.2 μm filtered solution in PBS, pH7.4.
Contact us for customized product form or formulation.
运输(Shipping)
This product is supplied and shipped with dry ice, please inquire the shipping cost.
存储(Storage)
Please avoid repeated freeze-thaw cycles.
This product is stable after storage at:
- 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.
质量管理控制体系(QMS)
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数据展示
电泳(SDS-PAGE)
Alpha-Synuclein monomer on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 90%.
活性(Bioactivity)-ThT Assay
Thioflavin T is a sensitive fluorescence reporter of fibrils formulation that binds to beta sheet-rich structures. The alpha synuclein pre-formed fibrils (Cat. No. ALN-H51H4) is able to induce the aggregation of alpha synuclein monomers (Cat. No. ALN-H52H8) (Routinely tested).
Protocol
电镜(Electron Microscope)
Transmission electron microscopy (TEM) of alpha-synuclein preformed fibrils (Cat. No. ALN-H51H4). Fibril structure is visible on negative stain TEM images of ALN-H51H4 (Routinely tested).
活性(Bioactivity)-CELL BASE
HEK293/Human Alpha-Synuclein (GFP) Stable Cell Line (Cat. No. CHEK-ATP085) were transduced with Human Alpha-Synuclein Pre-formed Fibrils, His Tag (Cat. No. ALN-H51H4) and Human Alpha-Synuclein, His Tag (Cat. No. ALN-H52H8) respectively. The fluorescence of GFP-Alpha-Synuclein (Green) and DAPI (Blue) were detected by confocal microscope. A. Lipo2000 transduction. B. Lipo2000 and Human Alpha-Synuclein, His Tag transduction. C. Lipo2000 and Human Alpha-Synuclein Pre-formed Fibrils, His Tag transduction. Scale bars, 50 μm (Routinely tested).
Protocol
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