RE: TUNEL Kits
T. Vincent Shankey (tshanke@bsd.medctr.luc.edu)
Thu, 24 Oct 1996 12:45:38 -0500 (CDT)
On Tue, 22 Oct 1996, Rakesh Nagarajan wrote:
>
> Hello,
> I would like to know which kit most people in your group use. The are
> two by Oncor, one a direct FITC conjugated dUTP and the other a
> Digoxigenin conjugated dUTP and an Ab to digoxigenin. Also, there is a
> kit available from Boehringer-Manheim.
>
> Thanks,
> Rakesh Nagarajan
> rakesh@milbrandt.wustl.edu
>
We have used the Oncor TUNEL kit, and our own "home brew" which follows
excellent suggestions from Zbigniew Darzynkiewicz - this includes the use
of BrUTP and an anti-BrdUr. In our experience, the sheep antibody and the
dUTP system in the original Oncor kit gave high background signals in
some types of human tissues. We have recently begun using a biotinylated
anti-BrdU (monoclonal Ab) provided to us by Phoenix Flow (an ironic name
for a company working on apoptosis-DEATH?), which, so far, has given
significantly lower levels of background signal (as compared with the two
antibody system we use - monoclonal anti-BrdU followed by biotinylated
goat anti-mouse Ig) for some of our "noisier" background types of human
tissues (prostate, testis). Overall, we have found systems using
incorporation of BrUTP with TdT significantly better than the dUTP
systems using polyclonal anti-dig. Phoenix may make this biotinylated
anti-BrdU kit available, with a little input from the field.
Vince Shankey, Loyola Univ. Med. Cntr.
CD ROM Vol 2 was produced by staff at the Purdue University Cytometry Laboratories
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