The TUNEL Assay
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Overview
One of the hallmarks of apoptosis is the nuclear DNA fragmentation by nucleases. These enzymes are activated by caspases, the family of proteins that execute the cell death program. TUNEL assay is a method that takes advantage of this feature to detect apoptotic cells. In this assay, an enzyme called terminal deoxynucleotidyl transferase catalyzes the addition of dUTP nucleotides to the free 3’ ends of fragmented DNA. By using dUTPs that are labeled with chemical tags that can produce fluorescence or color, apoptotic cells can be specifically identified.
JoVE’s video on the TUNEL assay begins by discussing how this technique can be used to detect apoptotic cells. We then go through a general protocol for performing TUNEL assays on tissue sections and visualizing the results using fluorescence microscopy. Finally, several applications of the assay to current research will be covered.
