ApopTag® Peroxidase in situ detection kits stains DNA strand breaks which in turn is detected by the TUNEL assay and subsequently visualised by light microscopy.
- Label apoptotic cells in research samples
- Modifies DNA fragments utilizing terminal deoxynucleotidyl transferase
- Chromogenic Detection
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
Principles of the procedure: Apoptotic cells are detected in situ by specific end labelling and detection of the DNA fragments produced by the apoptotic process. Residues of digoxigenin-nucleotide 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 single-stranded DNA. The incoorporated nucleotides form a random heteropolymer of digoxigenin-11-dUTP and dATP, in a ratio that has been optimized for antidigoxigenin 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.
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 digoxigenin-nucleotide 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 single-stranded DNA. The incorporated nucleotides form a random heteropolymer of digoxigenin-11-dUTP and dATP, in a ratio that has been optimized for antidigoxigenin 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.
