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Simple and Fast Rolling Circle Amplification-Based Detection of Topoisomerase 1 Activity in Crude Biological Samples
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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
A protocol for the sensitive and quantitative detection of topoisomerase 1 activity using the rolling circle enhanced enzyme activity detection assay is described. The method allows detection of topoisomerase 1 activity from purified components or cell/tissue extracts. This protocol has wide-ranging applications in any field involving detection of enzymatic activity.
