产品详情
分子别名(Synonym)
DDPAC, FTDP-17, MAPT, MSTD, MTBT1, Tau, PHF-tau, TAU
表达区间及表达系统(Source)
Human Tau-441 K18 (P301L) Pre-formed Fibrils Protein, Tag Free (TAU-H5113) is expressed from E. coli cells. It contains AA Gln 244 - Glu 372 (Accession # P10636-8 (P301L)).
Predicted N-terminus: Met
Request for sequence蛋白结构(Molecular Characterization)
This protein carries no "tag".
The protein has a calculated MW of 13.8 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)
>95% 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)
Tau-441 K18 (P301L) monomer on SDS-PAGE under reducing (R) condition. The gel was stained overnight with Coomassie Blue. The purity of the protein is greater than 95% (With Star Ribbon Pre-stained Protein Marker).
电镜(Electron Microscope)
Transmission electron microscopy (TEM) of Human Tau-441 K18 (P301L) Pre-formed Fibrils Protein, Tag Free (Cat. No. TAU-H5113). Fibril structure is visible on negative stain TEM images of TAU-H5113 (Routinely tested).
活性(Bioactivity)-CELL BASE
Human Tau-441 K18 (P301L) Pre-formed Fibrils Protein, Tag Free (Cat. No. TAU-H5113) induce tau phosphorylation and astrocyte activation in brain organoids (DIV100+, CIPO-BWL002K).
Protocol
活性(Bioactivity)-CELL BASE
HEK293/Human Tau (GFP) Stable Cell Line (Cat. No. CHEK-ATP087) were transduced with Human Tau-441 K18 (P301L) Pre-formed Fibrils Protein, Tag Free (Cat. No. TAU-H5113). The fluorescence of GFP-Tau (Green) were detected by confocal microscope. A. Lipo2000 transduction. B. Lipo2000 and Human Tau-441 K18 (P301L) Pre-formed Fibrils Protein , Tag Free (Cat. No. TAU-H5113) transduction. C. Score calculated from spot number/cell number. Both PEI and Lipo2000 can help tau-pFFs transfection and 4ug/ml is a better concentration when transfect cells in 24 well plate. Scale bars, 100 μm.
Protocol
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