Combination of TUNEL assay with immunohistochemistry for simultaneous detection of DNA fragmentation and oxidative cell damage.
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.
R&D Systems, Inc, Minneapolis, MN, USA.
Alexander E
AE
Kalyuzhny
D000080242:8-Hydroxy-2'-Deoxyguanosine; D000818:Animals; D002478:Cells, Cultured; D004249:DNA Damage; D053938:DNA Fragmentation; D003849:Deoxyguanosine / Q000031:analogs & derivatives; D005727:Ganglia, Spinal / Q000166:cytology; D007150:Immunohistochemistry; D020287:In Situ Nick-End Labeling / Q000379:methods*; D018384:Oxidative Stress; D016612:Paraffin Embedding; D051381:Rats; D017207:Rats, Sprague-Dawley; D013194:Staining and Labeling
D000080242:8-Hydroxy-2'-Deoxyguanosine; D003849:Deoxyguanosine