Related Experiment Video
Updated: Jun 6, 2026
Detection and Analysis of DNA Damage in Mouse Skeletal Muscle In Situ Using the TUNEL Method
Published on: December 16, 2014
Combination of TUNEL assay with immunohistochemistry for simultaneous detection of DNA fragmentation and oxidative
1R&D Systems, Inc, Minneapolis, MN, USA.
Abstract:
Oxidative cell damage causes disruption of DNA via formation of 8-hydroxy-2'-deoxyguanosine and can trigger apoptotic cell death. The cells damaged by oxidative stress can either become apoptotic, or recover. Therefore, it is helpful to employ a parallel assay that would confirm whether cells experiencing oxidative damage undergo apoptosis. Our paper describes the technique that combines immunohistochemical detection of 8-hydroxy-2'-deoxyguanosine with the TUNEL assay. This permits simultaneous detection of oxidative damage and apoptosis at a single-cell level. We have developed simple and reliable protocols which can be used with cultured cells and slide-mounted tissue sections. These techniques can be employed in research dealing with high-throughput drug screening, toxicology, and cancer.
Insights
This study presents a new method to detect oxidative DNA damage and apoptosis simultaneously in single cells. This combined assay aids research in drug screening, toxicology, and cancer studies.
Area of Science:
- Biochemistry
- Cell Biology
- Molecular Biology
Background:
- Oxidative cell damage, marked by 8-hydroxy-2'-deoxyguanosine, can lead to DNA disruption and apoptosis.
- Cells under oxidative stress may either undergo programmed cell death or recover, necessitating methods to distinguish these fates.
Purpose of the Study:
- To develop a parallel assay for confirming apoptosis in cells experiencing oxidative damage.
- To enable simultaneous detection of oxidative damage and apoptosis at the single-cell level.
Main Methods:
- Combining immunohistochemical detection of 8-hydroxy-2'-deoxyguanosine with the TUNEL assay.
- Developing simple and reliable protocols for cultured cells and tissue sections.
Main Results:
- Successful simultaneous detection of oxidative DNA damage and apoptosis in single cells.
- Validated protocols applicable to various biological samples.
Conclusions:
- The developed technique provides a reliable method for assessing oxidative stress and apoptosis concurrently.
- This approach is valuable for high-throughput drug screening, toxicology, and cancer research.
