[Histonet] RE: HIER - best retrieval with minimum tissue damage?

[C.M. van der Loos]

   Dear Nicola,

   In  our  experiments  we  saw  less  damage  by  HIER when applying an
   overnight   antigen   retrieval   at   60-70   C.   Especially   fatty
   tissues remain  in  tact nicely and do not tend to fall off. Using the
   ovenight  60-70C  procedure we saw that some antigens stain as good as
   with  15  min  boiling,  others  even more intense and others somewhat
   weaker.

   Hope this helps.

   Chris van der Loos, PhD
   Dept. of Pathology
   Academical Medical Center M2-230
   Meibergdreef 9
   NL-1105 AZ Amsterdam
   The Netherlands

   phone:  +31 20 5665631
   fax:    +31 20 6960389

   Date: Fri, 8 Jul 2005 10:41:21 +0100
   From: "Nicola Cragg" <n.cragg <@t> epistem.co.uk>
   Subject: [Histonet] HIER - best retrieval with minimum tissue damage?
   To: <Histonet <@t> lists.utsouthwestern.edu>
   Hello,
   I'm  a  big  fan  of microwaving in citrate buffer as a method of heat
   inducing epitope / antigen retrieval.  However, I do run into problems
   when  microwaving skin.  I've done a lot of IHC on skin and have found
   the  microwaving is great when staining antigens in the epidermis, but
   I do have problems if the antigens I want to look at are in the dermis
   and  require  microwaving  /  heat for retrieval.  The tissue is often
   damaged  and  the  morphology of the dermis is disrupted, so when I do
   see  evidence  of  specific staining it's difficult to be confident in
   it.
   Obviously,  when  optimising  antibodies,  I simultaneously attempt it
   without  antigen  retrieval  or  via other methods such as proteolytic
   digestion, but some antigens in t! he dermis