[Histonet] RE: permeabilization needed in IHC of viral protein?
Hi Carla,
Theoretically you don't need permeabilization. Since you have been
cutting sections, the cell interior is fully exposed to the
antibodies. On top of that you applied a protease step. My advise
would be testing the staining under similar conditions without and
with saponin 0.1% in all steps (including washing buffers). Again, to
my opinion a different staining pattern is not very likely.
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center
Amsterdam - The Netherlands
----- Original Message -----
From
Carla M Conway
Date
Wed, 02 Mar 2005 14:37:50 -0800
To
Histonet@lists.utsouthwestern.edu
Subject
[Histonet] permeabilization needed in IHC of viral protein?
Hello,
I am interested in using IHC to detect nucleocapsid (N) protein within
rhabdovirus. I have had success in detecting the glycoprotein on the
envelope surface, however the N protein is located within the virion.
Sections (FFPE) will be enzymatically digested with 0.05% protease XIV
in
TBS according to our standard protocol. Would a permeabilization agent
increase access to the interior proteins? Any suggestions/comments
will be
greatly appreciated.
Thank you,
Carla Conway
Western Fisheries Research Center
6505 NE 65th St
Seattle, WA 98115
ph: 206-526-6282 ext. 242
fax: 206-526-6654
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