RE: DAB question

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From:"Ana Moura" <mouraa@innogenex.com>
To:"Rebecca Hartley" <rebeccahartley@earthlink.net>, "'HistoNet Server'" <HistoNet@pathology.swmed.edu>
Reply-To:
Date:Wed, 28 Jul 1999 17:01:49 -0700
Content-Type:text/plain; charset="iso-8859-1"

  Hi everybody,
  Has anyone tried InnoGenex's Single-component DAB or AEC substrate (1 yr,
shelf life,@RT), instead of using the more cumbersome, and wasteful tablets?
  mouraa@innogenex.com
  www.innogenex.com

> -----Original Message-----
> From: Rebecca Hartley [mailto:rebeccahartley@earthlink.net]
> Sent: Friday, July 23, 1999 8:37 AM
> To: 'HistoNet Server'
> Cc: 'judy@playfair.utoronto.ca'
> Subject: RE: DAB question
>
>
> Judy (and anyone else interested!) -
>
> I still don't know why this problem with DAB loss with aqueous
> mounting is
> happening.  I had done this in the past with no problem, so ???  The only
> real difference from then to now was the DAB itself - this was the first
> time I've used the Sigma DAB tablets. But, I tried air drying the tissue
> and mounting with Cytoseal 60 (Stephens Scientific) and everything looks
> just fine.  I didn't use any alcohols, so you may be able to do
> this also.
>
> If anyone has any ideas on what's going on with this, I'd love to hear
> them!
>
> Rebecca Hartley
> Layton BioScience, Inc.
> Gilroy, CA
> rebeccahartley@earthlink.net
>
>
> Re:
> Date: 21 Jul 1999 09:25:44 -0500
> From: Judy Trogadis <judy@playfair.utoronto.ca>
> Subject: DAB question
>
> Dear Histonetters,
>
> We are staining cryosections of mouse embryos which have been frozen,
> sectioned and then fixed in paraformaldehyde. Immunostaining was carried
> out with the Vector Elite ABC system and the chromogen is DAB. After
> the DAB reaction is terminated with water, we don't dehydrate because
> we are also looking at retina which is easily damaged by alcohol,
> Therefore,we just use an aqueous mounting medium, either
> Ferrant's medium,
> or Aquamount.
>
> After coverslipping, the retinas look great. The mouse embryo DAB
> staining,
> however, quickly fades until it is barely visible, in a couple of hours.
> Any theories of how this could be happening?
>
> I should add that the retinas are fixed after removal from the animal,
> prior
> to freezing, but how could that make a difference to disolving the DAB
> precipitate?
>
>




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