Re: enhanced DAB kits, double staining

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From:Marek Malecki <>
To:Gayle Callis <>,
Content-Type:text/plain; format=flowed; charset=us-ascii

You can vary color and greatly enhance the signal by adding Ni or Co ions 
at the start of the DAB reaction.

At 8/27/00 Sunday04:11 AM+0000, you wrote:
>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

M. Malecki, M.D., Ph.D.
Director and Principal Investigator

Molecular Imaging Laboratories
University of California at San Diego
address: 1500 Suite Bonner Hall, 9500 Gilman Drive, La Jolla, CA 92093-0368
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