this is the problem with bone IHC.  for this reason i have converted most
hier procedures to enzyme ier.  i find that enzyme pretreatments are easier
to control than heat, you can use longer or shorter times and dilutions.
for most antibodies i use proteinase k for 5 min., or pepsin for 10 min. at
37dC.  i buy PK from DAKO and pepsin from Research Genetics.
Patsy Ruegg, retired
new email rueggp@earthlink.net 

-----Original Message-----
From: Coskran, Timothy M
To: Histonet (E-mail)
Sent: 8/24/01 9:24 AM
Subject: bone IHC

Hello,

Does anyone have any recommendations on how to keep bone sections on
your
slides through antigen retrieval steps involving heat?  We have tried
most
slide types and drying procedures that I'm aware of.  We have shown that
the
immunoreactivity we are looking for works best with steam citrate
retrieval.
However, the bones are the tissues of interest and we can never get past
the
point of retrieval.  All tissues are negative without retrieval.  We may
move into frozens, but are trying to salvage the formalin fixed paraffin
embedded tissue we have.

Thanks,
Tim Coskran

Discovery Pathology
PGRD
Pfizer
860-715-4558



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