Roche
In Situ Cell Death Detection Kit, POD
11684817910
The aim of the research is to detect apoptotic cells in formalin-fixed, paraffin-embedded (FFPE) human tissues. There are a few kits available on the market, but this one was recommended by a colleague. We wanted to compare the level of apoptosis in tonsil tissue among different groups of patients.
Immunohistochemsitry
FFPE tissue
The protocol is fairly standard for IHC, with the advantage of having a fluorescently labelled TdT which allows you to check the levels of labelling before adding the anti-fluorescein antibody Fab fragments (conjugated with horse-radish peroxidase) which catalyzes the substrate (DAB).
I would strongly recommend checking the levels of labelling (using a fluorescent microscope) before adding the anti-fluorescein antibody
The results at the first stage (before adding the anti-fluorescein antibody) looked good showing the "expected" low numbers of TUNEL-positive cells; however, after the converter-POD and substrate, we saw a lot more positive cells!
None
Easy to use
Discrepancy seen before and after POD/substrate
Interpretation of the data must be done with caution. I think imaging at the fluorescent stage is more reliable than post-conversion/substrate development.