[Histonet] Cleaved caspase 3

[Larry Woody]

We use the Cell Signaling Ab. with Citrate ph 6.0 and Dako Envision at 1:50 or 1.3ug/ml and it has been pretty good for the most part.
 
Larry A. Woody
Seattle, Wa.



----- Original Message ----
From: "McGinley,John" <John.McGinley <@t> ColoState.EDU>
To: "histonet <@t> lists.utsouthwestern.edu" <histonet <@t> lists.utsouthwestern.edu>
Sent: Monday, January 28, 2008 1:46:54 PM
Subject: [Histonet] Cleaved caspase 3

Hi,

Our lab recently purchased cleaved caspase 3 antibody (rabbit polyclonal) from Biocare for marking apoptosis, but the staining intensity is somewhat weak and does not appear to be marking the more classic examples of apoptotic cells that are typically identified via H&E. We tried both LSAB and Dako Envision on FFPE rat mammary tumors, but the staining patterns were similar. We've been using citrate buffer pH 6.0 for HIER, but we were wondering if anyone out there had better experience with other buffers like EDTA or maybe using enzyme pre-treatment in addition or instead of using HIER. Is this the best primary antibody of choice? Would the Cell Signaling antibody perform better on rat tissue?  Any input would be greatly appreciated. Thanks in advance.

Regards,

John McGinley
Cancer Prevention Laboratory
Colorado State University



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