Help with 2 color TUNEL

Stacy Ferguson sef at med.unc.edu
Thu Aug 8 06:25:59 EST 1996


In article <320940EF.125D at rex.re.uokhsc.edu>,
Kiley R. Prilliman <kprillim at rex.re.uokhsc.edu> wrote:
>I was wondering the exact same thing; could someone please be so kind as 
>to elaborate on this technique and what it is used for?
>
>Kiley R. Prilliman

It's a TdT-based system used to detect apoptotic cells. Since apoptotic
death results in the fragmentation of chromosomal DNA, this leaves
free DNA ends. The fragmentation can be detected _in situ_ by using
TdT to add detectable nucleotides to the free ends.

In the system I'm using, I'm adding digoxigenin labeled dUTP and adding
it to the ends with the TdT. Then the tissue sections are basically
treated like they would be for any other antibody staining by using an
anti-digoxigenin antibody (In my case, I'm using the antibody coupled
to horseradish peroxidase followed by development with a DAB substrate)
It's a nice clean assay that gives intense staining with virtually 
no background. 

If you want the original reference for the assay, email me. I'm at home
and don't have the volume and page numbers handy.

Stacy









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