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Biochemistry
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利用相扑融合策略生成国产、未加标签的杭丁顿蛋白 Exon1 单体和纤维
JoVE Journal
Biochemistry
This content is Free Access.
JoVE Journal
Biochemistry
Generation of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Using a SUMO Fusion Strategy
Please note that all translations are automatically generated.
Click here for the English version.
利用相扑融合策略生成国产、未加标签的杭丁顿蛋白 Exon1 单体和纤维
DOI:
10.3791/57506-v
•
11:22 min
•
June 27, 2018
•
Andreas Reif
,
Anass Chiki
,
Jonathan Ricci
,
Hilal A. Lashuel
1
Laboratory of Molecular and Chemical Biology of Neurodegeneration
,
Ecole Polytechnique Fédérale de Lausanne (EPFL)
Chapters
00:04
Title
00:53
Cell Lysis and Purification of His6-SUMO Httex1 Fusion Protein by Immobilized Metal Affinity Chromatography (IMAC)
02:56
Cleavage of the His6-SUMO-tag and HPLC Purification
06:06
Disaggregation and Resolubilization of Httex1 Proteins
07:53
Results: Characterization of Native, Untagged Huntingtin Exon1 Monomer and Fibrils Prepared via SUMO Fusion
09:52
Conclusion
Summary
Automatic Translation
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Automatic Translation
在这里, 我们提出了一个健壮和优化的协议, 以生产毫克数量的本地, 无标签单体和纤维的 exon1 的杭丁顿蛋白蛋白 (Httex1) 的基础上的瞬态融合的小泛素相关修饰剂 (相扑)。
Tags
Native Huntingtin Exon1
SUMO Fusion Strategy
SDS-PAGE Analysis
IMAC Purification
HPLC Purification
DTT
L-cysteine
Trifluoroacetic Acid
Aggregation-prone Proteins
Amyloid-beta
Alpha-synuclein
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