[Histonet] RE: permeabilization needed in IHC of viral protein?

From:"C.M. van der Loos"

   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 -----
   Carla M Conway 
   Wed, 02 Mar 2005 14:37:50 -0800
   [Histonet] permeabilization needed in IHC of viral protein?

   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
   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
Histonet mailing list

<< Previous Message | Next Message >>