Re: ER/PR inquiry

From:Richard Cartun

In order to respond to your e-mail I would like to know the following:

1.  Which ER/PR antibodies are you using?

2.  Are you using pretreatment?

3.  Which detection system are you using?

4.  Do you do your IHC manually or with automation?

R. Cartun

>>> K C  by way of Histonet  03/12/02 08:44AM >>>
I am seeking the experience of an ER/PR expert, or of
someone who reads ER and PR cases very frequently if
possible or someone who can get the info.  I have 2
questions or issues that need to be answered.

1.  Recently, I have been staining for ER and PR and
have noticed that in some cases, I get staining in the
cytoplasm of cells and negative nuclear staining in
the tumor.  In other cases, I get both nuclear
staining and cytoplasmic staining in the tumor.  Then
there are other cases that I get very clean staining
of tumor nuclei.  Sometimes, these circumstances occur
in one staining run.  All of this happens at the same
time that I get very clean negative controls.  My
question would be...are there ER and PR antigens that
sometimes reside in the cytoplasm and not in the
nucleus? and in what circumstances would this occur?

2.  In some cases of ER and PR, I notice that in a
particular run, I will get very bright staining of
nuclei on some slides, and on other slides in the same
run, I will get very faint staining.  My question is,
could this be biological (i.e. less antigenic sites
available), or is it more likely to be a prep problem
(retrieval, fixation)?  Please keep in mind that these
circumstance occur in one run--so I couldn't
understand how it could be retrieval.  I was very
interested to know this, because I know there are
scoring systems that take into account the intensity
of the staining, and if I try to correct for the
intensity by amplification, am I corrupting the actual
intensity of those slides?

Any help is extremely appreciated in this matter...


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