Teri,

The following instructions for creating positive control samples for
apoptosis are from the Intergen Company-ApopTag In Situ Apoptosis Detection
Kit :

1.  Pretreat section (they indicate any tissue will work) with DN Buffer
(30mM Trizma base-pH 7.2, 4mM MgCl2, 0.1mM DTT) at room temp for 5 min.

2.  Dissolve DNAse I in DN Buffer to final concentration of 1.0-0.1 ug/mL,
apply to sample and incubate 10 minutes at room temp.

3.  Rinse 5 times for 3 min. each with dH2O.

I have no experience using this protocol, but hope it works well for you!

Mandy




Mandy,

Yes, please do pass the information along.  The more I know about this, the
better!

Thanks,

Teri

-----Original Message-----
From: Mandy Annis [mailto:mandy_annis@usgs.gov]
Sent: Thursday, March 14, 2002 10:10 AM
To: Johnson, Teri
Subject: Re: control tissue for TUNEL



Teri,

My limited experience with apoptosis involved a kit from the Intergen
Company which supplied a rodent mammary gland (3-5 days post weaning) as a
positive control.  The instruction manual further suggests preparing a
positive control from any tissue by treating with DNase.  Full instructions
are supplied.  I would be happy to pass them on to you if this is of
interest.

Mandy

**********************************************************************
Mandy L. Annis
Columbia Environmental Research Center
4200 New Haven RD  Columbia, MO  65201

Phone:(573)-441-2940
Fax:(573)-876-1896
e-mail:mandy_annis@usgs.gov