[Histonet] weird brownish stuff in AEC slides

[Jan Shivers]

Is it brownish, as in a khaki color?  If so, that usually is a phenomenon
with AEC called metachromasia... a change in the precipitate color due to
molecules of chromogen being in too close in proximity.  Try diluting out
your primary antibody, so the Ag/Ab complexes aren't so close together.

Jan Shivers

----- Original Message ----- 
From: "SMITH,REBEKAH FELICIA" <rockbeki <@t> ufl.edu>
To: <histonet <@t> lists.utsouthwestern.edu>
Sent: Thursday, July 01, 2004 8:52 AM
Subject: [Histonet] weird brownish stuff in AEC slides


> I have yet another question, guys. I've been asking a lot of
> questions about blocking endogenous peroxidase,but now I have a
> new situation. I'm still using the ABC peroxidase method staining
> sheep placenta (formalin fixed, paraffin embedded) for eNOS and
> using AEC as a chromogen and gill's hematoxylin (blued with tap
> water and water with 2 or 3 drops of ammonia) as a counterstain.
> (btw, still blocking peroxidase with 30% peroxide in methanol,
> since my professors are away and haven't been able to order
> anything better) I have the lovely red and blue contrast in most
> parts of my slides, but I also am getting areas of brownish-black
> staining (kinda DAB colored, although I'm not using any DAB). I'm
> not exactly sure where the brown comes from this and whether its a
> problem with my AEC, my hematoxylin or something else. Anyone have
> any ideas? Thanks in advance, y'all have already been really
> helpful.
>
> --
> SMITH,REBEKAH FELICIA
> "You are a child of the universe, no less than the trees and the
> stars
> You have a right to be here and whether or not it is clear to you,
> no doubt the universe is unfolding as it should. Therefore be at
> peace with G-d, whatever you conceive Him to be. And whatever your
> labors and aspirations,in the noisy confusion of life, keep peace
> in your soul.-Max Ehrmann,"Desiderata"
>
>
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