[Histonet] EDTA decal

[Teri Johnson]

Hi Colleen,

I would say it's unusual, but not completely impossible that EDTA has interfered with your IHC.  We had that problem with demonstrating B-galactosidase in mouse bones. If we decalcified it in EDTA after whole mount staining in X-gal, the blue staining was removed. But if we decalcified it in formic acid, the stain was retained.

I think a few years ago, Biogenex had a room temperature antigen retrieval solution for acid decalcified bone (not the same as your situation, I know). But I think it was largely an alkaline solution you let the slides sit for maybe 30 minutes prior to staining. Might be worthwhile to try a different retrieval method for these and see what happens.

Otherwise, are you sure you are using a clone that reacts in mouse tissue? We use Rabbit monoclonal SP6.

Teri Johnson
Manager, Histology
GNF - San Diego, CA
858-332-4752