Double IHC staining question

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From:C & J Haley <>
To:HistoNet Server <>
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Hello Everyone:

I'm here to dip into the histo-wisdom pool again.  I've been doing some
double IHC staining.  This round of double staining involves two
antibodies requiring EDTA HIER.
Has anyone had experience with two EDTA requiring antibodies?  I've been
able to get staining, but the second antibody staining is not as
"crisp"  as I would like.  Any tips?  For those interested:  Yes, I am
actually doing two rounds of EDTA on the slides.  No, I'm not having any
section adhesion problems.  Yes, I've tried both alk. phos. and
peroxidase systems for the second round with the same results.

Thanks in advance,

Jane Haley
Barnes-Jewish Hospital
St. Louis, MO

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