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Cancer Research
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基于简单快速的滚动圆扩增检测粗生物样品中的拓扑异构酶1活性
JoVE Journal
Cancer Research
This content is Free Access.
JoVE Journal
Cancer Research
Simple and Fast Rolling Circle Amplification-Based Detection of Topoisomerase 1 Activity in Crude Biological Samples
Please note that all translations are automatically generated.
Click here for the English version.
基于简单快速的滚动圆扩增检测粗生物样品中的拓扑异构酶1活性
DOI:
10.3791/64484-v
•
10:13 min
•
December 02, 2022
•
Josephine Geertsen Keller
2
,
Karol Mizielinski
,
Kamilla Vandsø Petersen
2
,
Magnus Stougaard
3,4
,
Birgitta R. Knudsen
3
,
Cinzia Tesauro
1
Department of Molecular Biology and Genetics
,
Aarhus University
,
2
Department of Clinical Medicine
,
Aarhus University
,
3
VPCIR Biosciences ApS
,
4
Department of Pathology
,
Aarhus University Hospital
Chapters
00:04
Introduction
00:51
Preparation of Functionalized Slides
01:34
Generation of Closed Circular Substrates
02:16
Blocking of the Wellmaker and Hybridization of the Circles
03:48
Rolling Circle Amplification (RCA) and Visualization by Fluorescence Microscopy
05:54
Coupling of HRP-Conjugated Antibody to Rolling Circle Products and Visualization
07:57
Results: Detection of Topoisomerase 1 Activity Using the REEAD Assay
09:29
Conclusion
Summary
Automatic Translation
English (Original)
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Automatic Translation
描述了使用滚动圆增强酶活性检测测定法灵敏和定量检测拓扑异构酶1活性的方案。该方法允许从纯化的组分或细胞/组织提取物中检测拓扑异构酶1活性。该协议在涉及酶活性检测的任何领域具有广泛的应用。
Tags
Rolling Circle Amplification
Topoisomerase 1 Activity
Crude Biological Samples
Simple And Fast Detection
Anti-cancer
Anti-infectious Diseases
Primer Mix
Circular Substrates
Recombinant Topoisomerase 1
Cell Extract
SDS
Buffer Wash
Hybridization
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