[Histonet] anyone using Promega TUNEL assay?

[koellingr <@t> comcast.net]

Tyrone, 

I agree with Amos Brooks about the Chemicon kit and I agree with Jason Palmer about about Promega, especially in regards to smaller, controlled volumes to decease cost per slide.  In addition, I only used both those kits directions as starting points.  Took care of the proteinase digestion optimized to my standards for my controls in my lab and also emperically found a lesser amount (much less than suggested) of Tdt was preferable.  In fact, I could attribute false-positive staining to use of kit suggested amounts of Tdt.  Backing off on that concentration eliminated false positives while retaining strong pos staining. And made kit last longer. 



As for working up caspace-3, I'm sure you aware of the caveat that (1) brain especially can utilize non-canonical 

apoptosis pathways (caspace-3 independent pathways that leads to false negatives in the assay) and (2) there is now clear and convincing evidence that caspace-3 activity in brain has been linked to other things aside from apoptosis such as synaptic plasticity modulation and other non-apoptotic but normal brain functions. 



Ray Koelling 

PhenoPath Labs 

Seattle, WA  
----- Original Message ----- 
From: "Tyrone Genade" <tgenade <@t> gmail.com> 
To: "histonet" <histonet <@t> lists.utsouthwestern.edu> 
Sent: Saturday, July 11, 2009 6:11:53 AM GMT -08:00 US/Canada Pacific 
Subject: [Histonet] anyone using Promega TUNEL assay? 

Hello, 

For my PhD project my Prof and I are entertaining doing some TUNEL 
work on our brain sections in search of apoptotic nuclei but the kit 
is very expensive and would like to get some more information from 
some one who is using one of the kits and doesn't have a financial 
interest in selling us one. Locally, the Promega Kit has the highest 
tests to cost ratio. 

Anyone out there using this kit and willing to share their experience? 
-- 
Tyrone Genade 
http://tgenade.freeshell.org 
email: tgenade <@t> freeshell.org 
tel: +27-84-632-1925 (c) 
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