[Histonet] TUNEL

[Gayle Callis]

YFP is yellow fluorescent protein, is a chimera of Green fluorescent 
protein, the barrel shaped protein so popular these days.  I am not sure it 
can be referred to as an epitope, unless my definition of epitope is not 
complete or you are you are planning to detect the YFP with an antiYFP 
antibody.  Maybe I am missing something on the definition of 
epitope.  Being a chimera of GFP, it may be highly affected by fixation and 
other chemicals, temperatures, enzymes, even more so than eGFP itself, when 
used with TUNEL tissue preparations.   You can read up on working with GFP 
and YFP at the Clontech website - where you can access their manual on 
Living Colours.  I think there are now books written on the subject of 
using GFP and its chimeras.

I am not sure one can combine the use of YFP with TUNEL and get the best of 
both worlds, but a literature search in PUBMED may be helpful.

At 03:41 PM 1/3/2006, you wrote:
>I am going to begin some TUNEL experiments for the first time and have
>some basic questions.  First, how difficult is it to stain un-sectioned
>embryonic tissue?  The target cells are about 4-5 cell layers in from
>the surface.  Second, the target cells are transgenically labeled with
>YFP--will the YFP signal,or its epitope, be lost by the tissue prep for
>TUNEL?  Lastly, could I get some perspective on good/bad TUNEL kits?  I
>have found some of this topic in the archive, but this advice may have a
>shelf life.  Thanks in advance.
>
>Cordially,
>Noah Druckenbrod
>University of Wisconsin-Madison

Gayle Callis
Research Histopathology Supervisor
Veterinary Molecular Biology
Montana State University - Bozeman
PO Box 173610
Bozeman MT 59717-3610
406 994-6367
406 994-4303 (FAX)