[Histonet] Active caspase 3 staining of mouse small intestine

[Goodpaster, Tracy A]

We use the cleaved caspase-3 antibody from Biocare with good success.
Previously, we used the one from Cell Signalling, but switched after a
side by side comparison and cost evaluation.  We steam heat formalin
fixed, paraffin embedded sections for 15 minutes in Dako's pH10 antigen
retrieval buffer.  We then use an Avidin/biotin block and serum block
(15% goat + 5% human in antibody dilutent).  We incubate the antibody
for 90 minutes and detect it with a biotinylated goat anti-rabbit IgG at
1:400, the Vector elite RTU ABC and Dako DAB plus.  We get crisp
staining on mouse, dog and human tissue.  I hope this helps. 

-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Yu, Jian
Sent: Wednesday, February 01, 2006 9:56 AM
To: histonet <@t> lists.utsouthwestern.edu
Subject: [Histonet] Active caspase 3 staining of mouse small intestine

Thanks to several of you who have given me a lot of great suggestions on
how to get good cross sections of mouse small intestine   One of you
mentioned that you do Caspase 3 staining routinely with the small
intestine, sorry that I could not find your email anyone.  I have been
using the CAM1 Ab from BD, which works well for IF on frozen sections
but have a lot of background for IHC.  Unfortunately the frozen sections
do not have the greatest structures.  I would very much appreciate that
if you could share your experience with paraffin sections.

 

Thanks again!

********************************************************

Jian Yu, Ph. D.

University of Pittsburgh Cancer Institute

Hillman Cancer Center Research Pavilion

Office suite 2.26h, Laboratory 2.43

5117 Centre Avenue, Pittsburgh, PA 15213

 

Phone : 412-623-7786, (Lab) 412-623-3255

Fax:      412-623-7778

Email:   yuj2 <@t> upmc.edu

********************************************************

 

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