[Histonet] EDTA decal

[pruegg <@t> ihctech.net]

Good idea.  I have to soak the edta decaled tissues in an alkaline solution
inorder to restore enzyme histochemical staining for TRAP, might be the same
issue for some IHC.

Patsy Ruegg, HT(ASCP)QIHC
Ruegg IHC Consulting
40864 E. Arkansas Ave
Bennett, CO 80102
H 303-644-4538
C 720-281-5406
pruegghm <@t> hotmail.com 
rueggihcconsultingpr <@t> outlook.com



 

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-----Original Message-----
From: histonet-bounces <@t> lists.utsouthwestern.edu
[mailto:histonet-bounces <@t> lists.utsouthwestern.edu] On Behalf Of Teri Johnson
Sent: Friday, August 16, 2013 11:54 AM
To: 'cforster <@t> umn.edu'
Cc: histonet <@t> lists.utsouthwestern.edu
Subject: [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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