[Histonet] Decal/EDTA IHC

JColCLEFA <@t> aol.com JColCLEFA <@t> aol.com
Sat Sep 11 12:31:15 CDT 2004


In my lab, we use 10% neutral buffered formalin as a fixative for virtually 
all our material. We use a decal solution which contains both formalin and 
EDTA, and we use an "antigen retrieval" solution which contains EDTA for a 
significant number of proteins we wish to label. In my experience, certain primary 
antibodies will not work on material which was decalcified in a formalin/ formic 
acid/ sodium citrate/EDTA decal solution, but the vast majority will.   (Pax 
5 is one antibody which won't work for us on decalcified items)   My 
suggestion is to test all your variables before settling on your new EDTA decal. My 
assumption is that you run the risk of losing antigenicity on a few antibodies. 
To answer the question-- yes. Formalin will crosslink her antigenic sites, 
antigen (or epitope) retrieval procedures are successful at "decrosslinking" to 
enable labelling. My lab stains about 400 IHC slides daily, we formalin fix and 
subsequently perform antigen retrieval on virtually all our tissue. 40-50% of 
our work is on bone marrow specimens, and half of those are decalcified 
biopsies. JC--HT(QIHC)

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