DNA Fragmentation Detection Kit (30 slides)
AS20 4458 | The DNA Fragmentation Detection Kit can be used for the identification of apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue sections, or in preparations of single cell suspensions fixed on slides.
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Background | This DNA Fragmentation Detection Kit utilizes the fact that Terminal deoxynucleotidyl Transferase (TdT) binds to exposed 3’-OH ends of DNA fragments. During apoptosis, such DNA fragments are generated, which catalyzes the addition of biotin-labeled deoxynucleotides. A streptavidin-horseradish peroxidase (HRP) conjugate is used to detect biotinylated nucleotides, and diaminobenzidine (DAB) reacts with the HRP-labeled sample, generating a brown, insoluble substrate at the sites where DNA fragmentation has taken place. Counterstaining with Methyl Green allows the morphological evaluation and characterization of normal and apoptotic cells. The DNA Fragmentation Detection Kit can be used for the identification apoptotic nuclei in paraffin-embedded tissue sections, frozen tissue sections, or in preparations of single cell suspensions fixed on slides.
This kit contains the following reagents:
Proteinase K, a pH-stabilized Solution, 50 μl
TdT Equilibration Buffer, 4 ml
TdT Labeling Reaction Mix (mix of labeled and unlabeled nucleotides), 1.3 ml
TdT Enzyme (Terminal Deoxynucleotidyl Transferase), 40 μl
Stop Buffer, 4 ml
Block Buffer, 12 ml
Streptavidin-HRP Conjugate (25x concentrate), 150 μl
DAB Concentrate, 150 μl
DAB Reaction Buffer, 4 ml
Methyl Green Counterstain, 3.5 ml
This kit contains reagents for 30 tests.
Storage | Store kit at -20°C. For long-term storage, it is recommended to keep the TdT Enzyme, TdT Labeling Reaction Mix
and Streptavidin-HRP Conjugate (25x concentrate) frozen in aliquots. Use only the amount needed of these
components for each test, while leaving the remaining aliquots at -20°C. For short-term storage (up to 2 weeks),
the kit components can be kept at 4-8°C.
This kit should not be used beyond the expiration date.
Manual in pdf