dapi staining before secondary antibody

[fatboym]

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

[Dr.Stein]

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.

[Chris4]

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?

[Rene]

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