RE: bone IHC

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 

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


Does anyone have any recommendations on how to keep bone sections on
slides through antigen retrieval steps involving heat?  We have tried
slide types and drying procedures that I'm aware of.  We have shown that
immunoreactivity we are looking for works best with steam citrate
However, the bones are the tissues of interest and we can never get past
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.

Tim Coskran

Discovery Pathology

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