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Combination of TUNEL Assay with Immunohistochemistry for Simultaneous Detection of DNA Fragmentation and Oxidative Cell Damage

  • Alexander E. Kalyuzhny
Protocol
Part of the Methods in Molecular Biology book series (MIMB, volume 682)

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.

Key words

Oxidative stress, 8-hydroxy-2′-deoxyguanosine Immunohistochemistry TUNEL assay Apoptosis Cultured cells Tissue sections Double-labeling 

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Copyright information

© Springer Science+Business Media, LLC 2011

Authors and Affiliations

  • Alexander E. Kalyuzhny
    • 1
  1. 1.R&D Systems, IncMinneapolisUSA

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