enhanced DAB kits, double staining

<< Previous Message | Next Message >>
From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
Content-Type:text/plain; charset=us-ascii

We have been playing with DAB+ and AEC chromogens with double staining, but
haven't liked the color similarities (brown and red), a thing that is not
uncommon.  Am not sure black and red is so great either, but ------

***One thing the researcher is insisting on IS hematoxylin counterstaining,
no methyl green or nuclear fast red. 

What have people's experience with enhanced DAB kits, Zymed has one which
gives a dark brown color (DAB plus which is their DAB = enhancer solution)
or their DAB Black kit. Is the DAB highly sensitive, comparable to my
beloved DAKO DAB+.  

Have been disappointed in some of the other enhanced DAB kits or too
enthralled with some of the other HRP kits (messy background), that is
probably my control of the situation.

Hate giving up my beloved DAKO AEC+, unless I can postenhance this one,
some enhancing solutions are unbelievably pricey, others more reasonable.
Has anyone tried the Zymed enhancing solution (can be purchased separately)
with DAKO DAB or DAB+.  One company said theirs would work, but not willing
to pay the price.  

I refuse to weigh out, mess with DAB tablets in buffers - for some of the
metal enhancing methods, and this refusal includes Osmium tetroxide.

Just looking for a direction on what to try first, been in the Archives on

Guess the other question is what color combinations do you like best with
hematoxylin counterstain, and how much interference will hematoxylin have
with BCIP/NBT color. 

I could spend a fortune trying various companies products, but that is not
cost effective for research dollars. 

Still reading and sleuthing this one.

Gayle Callis
Veterinary Molecular Biology
Montana State University
Bozeman MT 59717-3610
406 994-4705
406 994-4303

<< Previous Message | Next Message >>