Apoptosis Detection
Apoptosis, or programmed cell death, is a key biochemical process whereby an organism regulates cell numbers and developmental pathways. This process has been shown to be a key factor in many diseases, including cancer, and is a target for research and drug discovery.

Qbiogene offers an expanding line of kits for in situ detection and analysis of one of the key characteristics of apoptosis - DNA fragmentation.

ApopTag^® Peroxidase in situ Oligo
Ligation (ISOL) Kit
While conventional labeling techniques are useful in detecting internucleosomal DNA cleavage, they cannot differentiate apoptotic from non-apoptotic DNA fragments. The new ApopTag^® ISOL kit uses a novel method to detect the type of DNA fragmentation that is specific to Apoptosis. It provides the best method for differentiating apoptotic cells from necrotic or transiently damaged cells.

During apoptosis, DNA cleavage results in blunt and short 3’ single base overhangs, as opposed to necrosis where the fragmented DNA will have 3’ and 5’ overhangs of varying length. The ApopTag^® ISOL method exploits this difference. Synthetic oligos labeled with biotin are selectively reacted with the specific types of genomic DNA ends characteristic of apoptotic cells. The reaction is mediated by T4 DNA ligase. Two different biotinylated, self annealing oligos are supplied in each kit. In the ISOL method, one of the oligos is attached to the fragmented DNA in situ, provided the DNA has a blunt end or a one base overhang on its 3’ end. Then a streptavidinperoxidase 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 which can be clearly visualized using brightfield microscopy.