[Histonet] RE: HIER - best retrieval with minimum tissue damage?
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
Hope this helps.
Chris van der Loos, PhD
Dept. of Pathology
Academical Medical Center M2-230
NL-1105 AZ Amsterdam
phone: +31 20 5665631
fax: +31 20 6960389
Date: Fri, 8 Jul 2005 10:41:21 +0100
From: "Nicola Cragg"
Subject: [Histonet] HIER - best retrieval with minimum tissue damage?
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
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
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