[Histonet] IHC anti-cleaved caspase 3

Liz Chlipala liz <@t> premierlab.com
Wed Sep 5 15:09:02 CDT 2007

I like the cell signaling antibody also, I tried biocare's but did not =
have much success with it.  The cell signaling antibody also works with =
pronase digestion, as well as the EDTA pH9 HIER.  We have even used it =
on bone sections with the pronase digestion. Works in multiple species, =
I have used it on human, rat, mouse, guinea pig, porcine and canine.  =
Our protocol is similar to Cheryl's.  It=92s a bit pricy as antibodies =
go, but I feel its worth it.  Good lot to lot consistency.  We have =
probably gone through about 5 different lots, with not changing the =
protocol at all.  I have a written protocol if anyone is interested.


Elizabeth A. Chlipala, BS, HTL(ASCP)QIHC
Premier Laboratory, LLC
P.O. Box 18592
Boulder, CO 80308
phone (303) 735-5001
fax (303) 735-3540
liz <@t> premierlab.com
Ship to Address:
Premier Laboratory, LLC
University of Colorado at Boulder
MCDB, Room A3B40
Boulder, CO 80309

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu =
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Cheryl =
Sent: Wednesday, September 05, 2007 2:01 PM
To: Thomas Pier
Cc: carl.hobbs <@t> kcl.ac.uk; histonet <@t> lists.utsouthwestern.edu
Subject: Re: [Histonet] IHC anti-cleaved caspase 3

I will second the Cell Signaling antibody - we have tried antibodies =
from Promega (which gave tons of background staining)...then via this =
site i was pointed to Cell Signaling's monoclonal which we tried; the =
polyclonal actually has been working very well with minimal background =
(we were getting lots of respiratory epithelium and endothelium lighting =
up in control animals). I have been using it in mice with this protocol:

primary antibody 60 minutes, dilution 1:125 Rabbit envision DAB plus

Hope this helps!

Cheryl Cross, DVM, Dipl. ACVP
University Corporation for Atmospheric Research College of Veterinary =
Medicine University of Tennessee Department of Pathology
2407 River Drive, Room A201
Knoxville, TN 37996-4542
(423) 967-2724
fax: 865-974-5616
ccross <@t> ucar.edu


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