ApopTag Peroxidase ISOL Apoptosis Detection Kit
- Novel labeling method clearly differentiates apoptotic from necrotic cells
- Minimizes background through high specificity of in situ labeling
- Reduces the incidence of false positives
The ApopTag Peroxidase ISOL Apoptosis Detection Kit is the newest member of the ApopTag family of Apoptosis Detection Kits. Like the original ApopTag kits, the ApopTag ISOL Kit detects DNA fragmentation by end-labeling the DNA. However, this kit uses a novel “In Situ Oligo Ligation (ISOL)” method to label the ends of DNA fragments that takes advantage of the prevalence of blunt-ends and 3’ singlebase overhangs characteristic only to apoptotic cells. This allows the ApopTag ISOL Kit to clearly differentiate cells undergoing apoptosis from cells undergoing necrosis.
The ApopTag ISOL Kit procedure attaches a specific biotinylated, synthetic oligonucleotide to the fragmented DNA in situ, provided that the DNA has a blunt end or a single base overhang on its 3’ end. Either Oligo A (3’ overhang) or Oligo B (blunt end) will bind to an exact complementary, doublestranded DNA end in a reaction catalyzed by T4 DNA Ligase. Then, a streptavidin-peroxidase conjugate is added that binds tightly to the biotin on the oligo. Lastly, the addition of DAB (diaminobenzidine), a chromogenic peroxidase substrate, causes a brown precipitate that can be clearly visualized using brightfield microscopy.
Detection of Apoptotic Cells in Rat Intestine Using the ApopTag ISOL Kit, Protocol One |
ApopTag ISOL Technology |
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