immunohistochemistry question

Dennis J. Young djyoung at ucsd.edu
Wed Apr 8 15:48:17 EST 1998


Rember trypsin is highly specific enzyme and won't "remove everything from
the surface"
It just clips a few peptide links! 
You should see only surface staining without the detergent. And you can ADD
extra cytokine for your positive control!


In article <Pine.SOL.3.91.980403105658.10642D-100000 at gsbs3.gs.uth.tmc.edu>,
"Amy E. Loercher" <aloer at gsbs3.gs.uth.tmc.edu> wrote:

:>        Howdy all--
:>
:>        We've been looking at cytokine production in peritoneal exudate 
:>cells by immunohistochemistry, and we've been seeing some cells stain 
:>positive for the presence of  certain cytokines, even though the specimen 
:>is negative for that cytokine transcript by RT-PCR. We think we're 
:>detecting cytokine that has bound to the cell's surface, but is not 
:>produced by the cell. R&D, the company that produces the MAb we're using, 
:>has said this is a possibility. Does anybody know a way to control for 
:>this? Right now, I'm thinking about either trypsinizing the cells prior 
:>to fixing to remove everything from the surface, or maybe running a 
:>sample without saponin and comparing the intensity of staining. Is there 
:>any reason why either of these would be better than the other? Thanks in 
:>advance.
:>
:>Amy Loercher
:>Dept of Gyn Oncology
:>MD Anderson Cancer Center



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