Cytokeratin (8/18)

From:James Mubiru <jamubiru@yahoo.com>

I recently did immunostaining for cytokeratin (8/18)
(NCL-5D3) on mouse prostate tissue. This antibody is
from Novacastra clone 5D3. It is a mouse monoclonal.
All my sections did not stain. I digested the tissue
with 0.4% trypsin for 10 minutes at room
temperature.The trypsin I use is the one from
Sigma(DPCC treated).  I know this antibody is supposed
to stain for glandular epithelium so the prostate
acinar cells should at least stain. The antibody
dilution I used was at 1:100 which is recommended by
the supplier. I incubated the primary antibody for 1
hour at room temperature. I also used the M.O.M kit
from vector since the primary antibody is a mouse
monoclonal and I am detecting in mouse tissue.
My question is if I try to use microwave treatment
with citrate buffer will I get a postive staining?. If
somebody has experience on this antibody on mice I
would appreciate the help. Thank you.
James Mubiru
UTHSCSA
San Antonio

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