AW: [Histonet] EDTA decal

[Gudrun Lang]

We experienced different IHC staining qualities on bone marrow, that was
decalcified with EDTA or formic acid. I remember, our CD38 antibody was
always very weak after EDTA and made no problems after formic acid.
I think it is very difficult to make an over-all-statement for all
antigens/all antibodies. 
Gudrun 



-----Ursprüngliche Nachricht-----
Von: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] Im Auftrag von Teri
Johnson
Gesendet: Freitag, 16. August 2013 19:54
An: 'cforster <@t> umn.edu'
Cc: histonet <@t> lists.utsouthwestern.edu
Betreff: [Histonet] EDTA decal

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

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