Re: [Histonet] Is TUNEL as waste of time?

From:John Kiernan

TUNEL and the related technique of in situ nick translation
are in situ end-labelling (ISEL) methods. They work by
adding labelled nucleotides to the fractures in broken 
DNA molecules. Many cells peg out quietly (apoptosis),
either because they are no longer needed (normal) or 
because of disease. In a cell that's about to die, 
enzymes become active that cut DNA into short fragments.
The cut ends provide binding sites for the labelled 
nucleotides used in ISEL methods.
Satoshi Akima wrote:
> Dear all,
> I am looking into the Roche TUNEL kit and have been getting various
> opinions. Some say it is easy and reviewers will ask for a TUNEL stain
> in preference to other methods such as staining for caspase-3. However
> many people find that they get non-specific staining and even where
> things seem to have gone well you get a great many cells that seem
> absolutely healthy staining up positive.
> Do people think that TUNEL is a sham technique that ivory tower
> reviewers who never get their hands dirty in the lab ask for, never
> realising how hopeless the technique is? Or is it still a genuinely
> useful tool?
> Toshi Akima
> PhD Student
> Centre for Transplantation and Renal Research
> Westmead Millenium Institute
> Sydney, Australia
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