[Histonet] RE: promega tunel

[PALMER Jason (SVHM)]

I have used a Promega Dead-End kit for a few years now and am happy enough with the results - get consistent TUNEL labelling with a biotin / DAB kit from run to run, and get away with using smaller volumes than they suggest, with coverslips for the reaction mix incubation.  The only thing is (and I think this is true for TUNEL in general) that interpretation can be tricky as there seem to be a lot of false positive cells, especially the longer you do your proteinase K digestion for.  (Negative controls are clean.) So you may need to confirm what you get morphologically (not always easy, depending on your experience and the cells you are looking at) or by some other method - I am trying to work up cleaved caspase 3 immunostaining at the moment for just this purpose.

Jason Palmer
Histology Laboratory Coordinator
Bernard O'Brien Institute
42 Fitzroy St, Fitzroy Victoria 3065
Australia
tel +61 3 9288 4018
fax +61 3 9416 0926
email: jason.palmer <@t> svhm.org.au

Message: 9
Date: Sat, 11 Jul 2009 15:11:53 +0200
From: Tyrone Genade <tgenade <@t> gmail.com>
Subject: [Histonet] anyone using Promega TUNEL assay?
To: histonet <histonet <@t> lists.utsouthwestern.edu>
Message-ID:
        <a801af370907110611t3c1b3518qd6ba156f50dc6e24 <@t> mail.gmail.com>
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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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End of Histonet Digest, Vol 68, Issue 11
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