[Histonet] decal and ihc

louise renton louise.renton <@t> gmail.com
Wed Jun 14 02:46:55 CDT 2006

dear all,
on pain of revisiting stuff that may or may not have been addressed before-

I have just done some immuno on FFPE tissue that had been decalcified
in a decal mix of 10% formic , 10% HCl and 1% Na Citrate. Duration in
acidic soln is unknown, as these blocks date from before my employment
here. On performing immuno, I found that the staining is "muddy" and
non specific. Is this the usual consequence of decalcification or did
I unwittingly do something wrong?. The antibody used has been
optimised on normal FFPE tissue, and worked fine, with clean and crisp
staining. I was always under the impression that decalcification
destroyed antigenicity and thus caused less staining rather than

Having given this scenario, is there anything I can try to improve
matters? (poly rabbit antibody incubated overnight @4deg C. ABC
amplification and DAB chromogen. No pretreatment performed, washes in
PBS with tween)
 Best regards
Louise Renton
Bone Research Unit
University of the Witwatersrand
South Africa
"There are nights when the wolves are silent and only the moon howls".
George Carlin
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