[Histonet] Re: dissolving DAB

[David A. Wright]

Hi folks

I'm at home without my notes, but I remember John Kiernan
saying somewhere (Histonet, the Book?) that DAB dissolves
better in pure water.  Since then that's what I've done -
dissolve in deionized water at 1mg/ml, vortex thoroughly, then
bring up to rxn volume in Tris, pH 7.6.  Works a treat!

I stain thick (40um) sections, so I also like to preincubate
in my DAB solution plus any intensifiers for 8-10 min to allow
them to penetrate, then start the reaction by adding the
peroxide later - it's a tiny molecule and gets everywhere very
quickly.

-david
--
David A. Wright, PhD
Section of Neurosurgery
University of Chicago 
================
ORIGINAL POST:
Histonet Digest, Vol 57, Issue 41: Message: 2
Date: Fri, 29 Aug 2008 10:06:55 -0700 (PDT)
From: Kim Merriam <kmerriam2003 <@t> yahoo.com>
Subject: [Histonet] DAB: old-fashioned style!
To: Histonet <histonet <@t> lists.utsouthwestern.edu>

Hi All,

It's been many moons since I had to make DAB by hand and I
have lost my recipe.  Can anyone let me know how to make DAB
from powder and H2O2?

Thanks,
Kim
�Kim Merriam, MA, HT(ASCP)QIHC
Cambridge, MA