Re: elution of antibody complex for double IHC staining

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From:John Bliss <>
To:Gayle Callis <>
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Dear Gayle:

Zymed sells the Histostain-DS kit (cat. no. 95-9999) for doublestaining any
combination of mouse, rabbit, rat, or guinea pig primary antibodies on human
tissues. This kit uses a double staining enhancer (comparable to the
doublestain block) that is sold separately (cat. no. 50-056).


John Bliss
Zymed Laboratories, Inc.

Gayle Callis wrote:

> AFter visiting with DAKO tech services this am, it was pointed out their
> their Envision double staining kit uses a special "Doublestain block" to
> remove any residual immune complex AFTER the DAB step, in order to prevent
> stearic hinderance and allow the next antibody to bind to its antigen
> without another immune complex in the way.  This treatment was said to be
> handy for co-localization of two markers on a cell (double IHC)
> This is part of a kit, but it is also mentioned later on in catalog, 0.03N
> HCl has also been used for this purpose, which as an added bonus, blocks
> all endog alk phos thereby eliminating Levamisole step for the second alk
> phos substrate.
> Has anyone tried using acetic acid, which at 15% will block gut endog alk
> phos.
> If acetic acid can is successful, at what concentration was successful, and
> timing (probably in the 3 - 5 min range).  I am not too enthralled with
> HCl, and a milder acid might work???
> Unfortunately, they do not sell this handy doublestain blocker as a
> separate reagent and their mixture is proprietary.  DAKO is listening I
> hope as it would be nice as a single reagent i.e. same as your other
> separate blocking reagents!
> Gayle Callis
> Veterinary Molecular Biology
> Montana State University
> Bozeman MT 59717-3610
> 406 994-4705
> 406 994-4303

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org:Zymed Laboratories
title:Director of Marketing
note;quoted-printable:458 Carlton Ct, South San Francisco, CA, 94618 =0D=0ATel: 650-871-4494 FAX: 650-871-4499
fn:John Bliss


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