apoptosis essay in dorsal root ganglion tissue

Jason L. Eriksen jerikse at news.luc.edu
Wed Nov 8 13:04:40 EST 1995


karl-heinz blenk (Karl at blenk.deceiver.org) wrote:
: 1) I am looking for a marker to detect apoptosis in dorsal root ganglion 
: cells using light microscopy.
: I con faintly remember having heard something about Acrydyl-orange.
: Does anybody has any experience with this substance or any other 
: suggestions to offer?

Karl-Heinz,

I'm also sending you an e-mail about this, but for the benefit of everyone
else:

You didn't say the conditions of your experiments, but if you are planning
to fix the tissue, I'd recommend performing TUNEL or ISEL techniques for
in situ staining.  TUNEL is a fairly simply procedure which relies upon
DNA fragmentation in apoptotic cells as a marker.  TdT enzyme appends
base analogs to the 3OH end of DNA fragments.  Next, you add a primary
antibody that tags the analogs.

It's a clever, simple technique that works pretty well (but is also fairly
expensive).  

ISEL is a variant of TUNEL.

You can get a TUNEL stain kit, called APOPTAG, from Oncor (USA).  Ask for 
the Peroxidase kit, which is a light stain.  Kit cost is $300, which allows
you to do up to 50 slides (but probably 40 is more realistic).  You'll also
need to develop a positive control protocol, requiring a bit of fooling around
with times of exposure. I have  a protocol if you're interested.

Jason Eriksen



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