Re: [Histonet] Melanin bleaching and IHC-P

From:"Mark Tarango"

Hi Stephan,

Yep, you don't have to bleach it because this is immunofloresece.

People bleach because melanin pigment can look a lot like the chromogen
DAB.  I think it's better to just do it in red, if it were light microscopy


On 8/11/08,  wrote:
> 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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