RE: [Histonet] IHC anti-cleaved caspase 3

From:"Liz Chlipala"



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.  Itís 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.

Liz

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

-----Original Message-----
From: histonet-bounces@lists.utsouthwestern.edu [mailto:histonet-bounces@lists.utsouthwestern.edu] On Behalf Of Cheryl Cross
Sent: Wednesday, September 05, 2007 2:01 PM
To: Thomas Pier
Cc: carl.hobbs@kcl.ac.uk; histonet@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:

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

Hope this helps!

Cheryl Cross, DVM, Dipl. ACVP
Researcher
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@ucar.edu




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