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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
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JoVE Journal Cancer Research
Simple and Fast Rolling Circle Amplification-Based Detection of Topoisomerase 1 Activity in Crude Biological Samples
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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

DOI:
10.3791/64484-v

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10:13 min

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December 02, 2022

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, , , , ,
1Department of Molecular Biology and Genetics,Aarhus University, 2Department of Clinical Medicine,Aarhus University, 3VPCIR Biosciences ApS, 4Department of Pathology,Aarhus University Hospital

Chapters

  • 00:04Introduction
  • 00:51Preparation of Functionalized Slides
  • 01:34Generation of Closed Circular Substrates
  • 02:16Blocking of the Wellmaker and Hybridization of the Circles
  • 03:48Rolling Circle Amplification (RCA) and Visualization by Fluorescence Microscopy
  • 05:54Coupling of HRP-Conjugated Antibody to Rolling Circle Products and Visualization
  • 07:57Results: Detection of Topoisomerase 1 Activity Using the REEAD Assay
  • 09:29Conclusion

Summary

Automatic Translation

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 AmplificationTopoisomerase 1 ActivityCrude Biological SamplesSimple And Fast DetectionAnti-cancerAnti-infectious DiseasesPrimer MixCircular SubstratesRecombinant Topoisomerase 1Cell ExtractSDSBuffer WashHybridization

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