peritoneal cells

[Ekaterina E. Zueva]

        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


What kind of slides do you use? The cells from peritoneal fluid - there
will be cyto- but not histo chemistry.
Yekaterina Zueva