ApopTag^® Peroxidase in situ Detection Kits
TUNEL kit for use with light microscopy

ApopTag^® Peroxidase in situ Detection Kits detect apoptotic cells by immuno-peroxidase detection of digoxigenin-labeled genomic DNA in thin sections of fixed tissue. The ApopTag^® Peroxidase Plus kit includes positive control slides to aid identification of apoptotic cells.

Principles of the procedure: Apoptotic cells are detected in situ by specific end labeling and detection of the DNA fragments produced by the apoptotic process. Residues of digoxigeninnucleotide are catalytically added to the DNA by terminal
deoxyribonucleotidyl transferase (TdT), an enzyme which catalyzes a template-independent addition of deoxyribonucleotide triphosphate to the 3’-OH ends of double- or singlestranded DNA. The incorporated nucleotides form a random heteropolymer of digoxigenin-11-dUTP and dATP, in a ratio that has been optimized for anti-digoxigenin antibody binding.
The anti-digoxigenin antibody fragment carries a conjugated reporter enzyme (peroxidase) to the reaction site. The localized peroxidase enzyme then catalytically generates an intense signal from a chromogenic substrate. This system allows for
sensitive and specific staining of the very high concentration of 3’-OH ends that are localized in apoptotic bodies.

Kit Components: Equilibration Buffer, Reaction Buffer, TdT Enzyme, Stop/Wash Buffer, Plastic Coverslips, and Anti-Digoxigenin-Peroxidase