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Counterstain for AEC AND BCIP/NBT

Counterstain for AEC AND BCIP/NBT - Protocols and Methods Forum

Counterstain for AEC AND BCIP/NBT - Post Any Protocol, Method, Technique, Procedure or Tips / Troubleshooting for any Molecular Biology Technique.


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  #1  
Old 08-27-2008, 08:30 AM
Bluescript
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Default Counterstain for AEC AND BCIP/NBT



Hi,
I want to do immunohistochemistry detecting two targets at the same
time. For various reasons I am going to use AEC as substrate for HRP
and BCIP/NBT as AP substrate. Now I am looking for a good
counterstain. Because of the AEC the counterstain has to work with an
aqueous embedding medium.
Has anybody a good idea? Thank you very much for your help in advance,
Sven
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  #2  
Old 08-27-2008, 07:10 PM
Dr Engelbert Buxbaum
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Default Counterstain for AEC AND BCIP/NBT

Am 27.08.2008, 04:30 Uhr, schrieb Bluescript <[Only registered users see links. ]>:


Intercalating DNA stains are commonly used to mark nuclei: bisbenzimide,
DAPI, Propidium iodide and a host of proprietary dyes (check the Molecular
Probes catalogue).
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Old 08-28-2008, 07:06 AM
Bluescript
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Default Counterstain for AEC AND BCIP/NBT

On 27 Aug., 21:10, "Dr Engelbert Buxbaum"
<[Only registered users see links. ]> wrote:

Thank you very much for your suggestions. Maybe I forgot to mention
that I am looking for a chromogenic counterstain that can be
visualized by light microscopy!
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  #4  
Old 09-05-2008, 07:11 PM
Dr Engelbert Buxbaum
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Default Counterstain for AEC AND BCIP/NBT

Am 28.08.2008, 03:06 Uhr, schrieb Bluescript <[Only registered users see links. ]>:



Classical nuclear stains like haematoxilin, methylgreen, nuclear fast red
should work, perhaps you'll have to apply them before immunostaining.
Methylenblue-eosinate (Giemsa, Wright, Papenheim...) would give blue
nuclei and pink cytosol, you'll have to check whether the contrast with
the immunostaining is sufficient. Perhaps switch to
Diaminobenzidine/Nickel for HRP detection (black, chemically quite inert).
But overall, I'd recommend swithching to fluorescence detection, check
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