Re: [Histonet] Immunoperoxidase Double staining protocol

From:Rene J Buesa

Generally speaking you will have to run the whole protocol for one of the Abs and use one chromogen (of your selection). After that, since you already did HIER and blocked the internal peroxidase, you will treat the section with the second Ab, will link/detect it and use another chromogen that ideally should have a different color.
  The first chromogen could be regular DAB and for the second you can use also DAB but with incorporated nickel that will give a deep/blue purple color, in contrast with the brown/reddish normal color of the DAB.
  The thing is that you don't need any special protocol, just your regular IHC protocol for FFPE tissue, run twice with two different chromogens.
  René J.

Reuel Cornelia  wrote:
  I was asked to do an immunoperoxidase double staining on CD61 ,Cd42, pf4 with an unknown mouse antibody that will work on platelets. Can somebody help me or any protocol available that I can refer to. I am used of IF double stianing but not Immunoperoxidase. Thank you.

Reuel Cornelia, BS MT, AMT
Cellular Pathology
Texas Scottish Rite Hospital for Children
2222 Welborn Street
Dallas, TX 75219
Tel: 214-559-7766
fax: 214-559-7768

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