c-Fos DAB staining fades?

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kerribridgeslab
kerribridgeslab's picture
c-Fos DAB staining fades?

I have run IHC on rat brain sections fixed in 4% PFA. After using an ABC detection kit followed by DAB staining (enhanced with NiCl) I've found that the staining in my sections has faded over the weekend. (They were a very dark color on Thursday). They were left out in the light, although I wouldn't think this type of staining is very susceptible to photobleaching. Any thoughts as to what caused the fading?
Thank you

Fraser Moss
Fraser Moss's picture
DAB is light sensitive, so if

DAB is light sensitive, so if your samples were left out and in particular, close to a fluorescent light source you may well indeed see significant fading.

Also DAB is sensitive to the pH of the mounting medium. Acidic or basic pH's can accelerate fading - check the neutrality of the mounting medium that you are using.