dapi staining before secondary antibody

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    • #1706
      fatboym
      Participant

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

    • #28740
      Dr.Stein
      Participant

      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.

    • #28749
      Chris4
      Participant

      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?

    • #28814
      Rene
      Participant

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

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