S'identifier

JoVE Journal

Cancer Research

This content is Free Access.

live

Speed

×

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

2.3K Views

10:13 min

December 2nd, 2022

December 2nd, 2022

2,292 Views

0:04

Introduction

0:51

Preparation of Functionalized Slides

1:34

Generation of Closed Circular Substrates

2:16

Blocking of the Wellmaker and Hybridization of the Circles

3:48

Rolling Circle Amplification (RCA) and Visualization by Fluorescence Microscopy

5:54

Coupling of HRP-Conjugated Antibody to Rolling Circle Products and Visualization

7:57

Results: Detection of Topoisomerase 1 Activity Using the REEAD Assay

9:29

Conclusion

Transcription

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.

Nous utilisons des cookies afin d'améliorer votre expérience sur notre site web.

En continuant à utiliser notre site ou en cliquant sur le bouton ''continuer'', vous acceptez l'utilisation de cookies.