[Histonet] decal and ihc

[louise renton]

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
more??!!

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
Johannesburg
South Africa
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