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Methods Mol Biol. 2011;682:15-27. doi: 10.1007/978-1-60327-409-8_2.

Combination of TUNEL assay with immunohistochemistry for simultaneous detection of DNA fragmentation and oxidative cell damage.

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1
R&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.

PMID:
21057917
DOI:
10.1007/978-1-60327-409-8_2
[Indexed for MEDLINE]
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