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When cells undergo apoptosis, they activate endonuclease enzymes that cut genomic DNA between nucleosomes. DNA ladder of 180-200 bp can be detected by electrophoresis after extraction of DNA during apoptosis.
TUNEL (TdT mediated dUTP Nick End Labeling) Cell apoptosis Assay Kit (FITC) can be used to detect the nuclear DNA breakage in tissue cells during late apoptotic stage. The principle is that FITC-12-dUTP is inserted into the 3'-hydroxyl (3'-OH) terminal of genomic DNA exposed during DNA break under the action of Terminal Deoxynucleotidyl Transferase (TdT). This can be detected by fluorescence microscopy or flow cytometry.
This kit optimizes the labeling reaction by using FITC-12-dUTP and unlabeled dNTP at the optimal ratio for nucleotide incorporation at the 3' -OH terminal, resulting in a longer "labeling tail" at the end of the same broken DNA fragment. The “labeling tail” reduces the steric hindrance of labeled groups on adjacent dNTP, increases the number of fluorescent-groups on each broken fragment, and reduces the aggregation and quenching that may be caused by adjacent fluorescent-groups, thus improving detection sensitivity and reducing non-specific reactions.
This kit has a wide range of applications. It can be used to detect apoptosis in frozen or paraffin sections, as well as in culture adherent cells or suspension cells situation.
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