RE: [Histonet] Active caspase 3 staining of mouse small intestine

From:"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-----
[] On Behalf Of Yu, Jian
Sent: Wednesday, February 01, 2006 9:56 AM
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




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