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 -
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.
|ApopTag® ISOL Kit
||NEW TECHNIQUE based on ligation of labeled hairpin oligo to apoptosis- specific DNA fragments. Provides best possible differentiation
of apoptosis from necrosis.
||TUNEL method, DIG labeled dUTP, detection with Ab-peroxidase conjugate
||TUNEL method, DIG labeled dUTP, detection with Ab-fluorescein conjugate
|ApopTag® Fluorescein Direct
||TUNEL method dUTP direct-labeled with fluorescein
|S7200-KIT ApopTag® (ISOL) Kit 40 tests