[Histonet] Background problems with frozen liver IHC

[James S.]

Hi All,

I do a lot of fluorescence labelling of frozen mouse tissue (mostly spleens) but recently I've been trying to do IHC staining of various mouse (and human) tissues. I'm having particular trouble with liver samples with high brown background in the hepatocytes - the vascular areas are completely clean. I've been using Vector Labs avidin/biotin blocking system and their ABC detection method along with DAB (Dako) and various biotinylated secondary antibodies. There is background from the secondary antibodies and to a lesser extent from the ABC.

Any tips?

Thanks
Sonya