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TUNEL Assay

QC Qingxia Chen
JY Jingjia Yan
WX Wenji Xie
WX Wenqin Xie
ML Meijun Li
YY Yanle Ye
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Apoptosis-related DNA fragmentation was analyzed by terminal deoxynucleotidyl transferase-mediated deoxynucleotidyl triphosphate in situ nick-end labeling (TUNEL) staining using in situ apoptotic cell death detection kit (Roche Applied BioSciences) according to the manufacturer’s instructions. The cells cultured on the coverslips were washed with PBS and fixed with ice-cold 1% paraformaldehyde. Subsequently, the terminal deoxynucleotidyl transferase (a template-independent polymerase) was employed to incorporate nucleotides at the DNA cleavage site. Following that, nuclei were stained with TO-PRO®-3 and fluorescence images were taken on three different fields of view of each coverslip using a confocal microscope. The apoptotic index was calculated as a percentage according to the following formula: TUNEL positive cell number/total cell nucleus.

Copyright and License information:  ©2020 Chen, Yan, Xie, Xie, Li and Ye.
This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

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