DNA-based assay of apoptosis needed

Julie Roberts jroberts at rascal.med.harvard.edu
Wed Feb 5 09:22:11 EST 1997


Jerry Kropp wrote:
> 
> Anyone out there know of a reference or protocol to assay for apoptosis,
> DNA-wise? Someone here mentioned a procedure acronymed TUNEL, but that's
> all he knew.
> Apologies if this appears more than once, my news server(s) and I are estranged.
> Thanks in advance.
> Jerry
> 
> --
> "If it ain't broke, it needs more features"-plagerized


I have used the Boehringer Mannheim TUNEL kit though not for
quantification. This kit enzymatically labels strand breaks with
flourescent dUTP, you can also get a TUNEL kit that utilizes alk phos as
well. It takes about half a day to do the assay. I did mostly microscopy
fixes and didn't have much sucess with the flow cytometry although I was
using primary macrophages and this could have been part of the proble.
There is another kit by clonetech which utilizes Annexin V that I had
really nice results with, again I'm not sure if this is quantitative.
Someone else in the lab was doing a DNA laddering assay but I don't
think that is quantitative either.

Julie

jroberts at rascal.med.harvard.edu



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