elution of antibody complex for double IHC staining

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From:Gayle Callis <uvsgc@msu.oscs.montana.edu>
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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

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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