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Your Position: Home > Protein > Alpha-Synuclein > ALN-H51H4

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

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  • Synonym
    SNCA, NACP, PARK1, alpha-Synuclein
  • 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
  • Molecular Characterization
    Alpha-Synuclein Structure

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

    The protein has a calculated MW of 16.3 kDa.

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

    >90% as determined by SDS-PAGE.

  • Formulation

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

    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 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.
SDS-PAGE
Alpha-Synuclein 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%.

Bioactivity-ThT Assay
 Alpha-Synuclein THT

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).

Electron Microscope
 Alpha-Synuclein 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).

Bioactivity-CELL BASE
 Alpha-Synuclein CELL

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).

  • Background
    Alpha-synuclein is a neuronal protein that plays several roles in synaptic activity such as regulation of synaptic vesicle trafficking and subsequent neurotransmitter release. It acts also as a molecular chaperone in its multimeric membrane-bound state, assisting in the folding of synaptic fusion components called SREs (Soluble NSF Attachment Protein REceptors) at presynaptic plasma membrane in conjunction with cysteine string protein-alpha/DJC5. Abnormalities in alpha-synuclein are implicated in the pathogenesis of Parkinson's disease (PD). Alpha-synuclein is present in Lewy-bodies, the neuropathological hallmark of PD, and the protein and its aggregation have been widely linked to neurotoxic pathways that ultimately lead to neurodegeneration.
  • Clinical and Translational Updates

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  • Latest Research Phase:Phase 3 Clinical

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