dapi staining before secondary antibody

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fatboym
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dapi staining before secondary antibody

Post by fatboym » Fri Aug 19, 2005 9:20 am

I have stained with dapi before adding the secondary antibody. can I added now or remove the dapi and how? :oops:

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Dr.Stein
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Post by Dr.Stein » Sun Aug 21, 2005 11:57 pm

dapi? You mean DAP, the chromogen substance? In my protocol, I add DAP after applying secondary antibody because it is a marker to see whether there is bond between antigen and primary antibody (monoclonal), which can be detect by bond between that primary antibody and secondary antibody (polyclonal). Thus I think, the secondary antibody must be placed before the chromogen. The principle of IHC or ICC is similar to ELISA's.
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Chris4
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Post by Chris4 » Mon Aug 22, 2005 8:26 am

No they mean Dapi. Stains the nucleus blue. I don't know whether you can remove it. I don't think so.
Have you done the experiment yet? did it work?
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Rene
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Post by Rene » Wed Aug 24, 2005 11:58 am

You can add the secondary antibody after staining with Dapi, no problem at all.

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