[Histonet] Melanin bleaching and IHC-P

From:hyters@onid.orst.edu



Hello,

I am attempting to perform IHC-P on heavily pigmented melanocytic  
growths using flourescent antibodies.  I have tried using 10% H2O2  
both overnite at room temperature and over two hours at 60 degrees.   
The bleaching is decent but tissue integrity suffers greatly.

I have not tried the potassium permanganate/oxalic acid method yet  
(although at this point I will have to try it) but I had a question  
about the necessity of bleaching as I am not using chromagen-based  
antibodies.  The bleaching does not remove the pigment (correct?) so  
it is not a question of primary antibody accessibility but of color  
differentiation right?  Since I am using flouresence microscopy is  
bleaching the growths a necessary step?

Thanks for your time,
Stephen

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