Protocol Online logo
Top : New Forum Archives (2009-): : Immunology

DAB staining and GFP - (Feb/15/2012 )

I'm just about to begin injecting mouse brains with a GFP-tagged lentivirus. After sacrificing, the brains will be sectioned on a microtome and immunohistochemistry will be performed on the free floating sections. I was wondering if doing DAB staining affects the GFP fluorescence at all? I will want to see co-localisation of different markers with GFP-positive cells so would it be better to stain the cells using fluorescently tagged secondary antibodies or would the GFP expression still be present after doing the DAB reaction?



In our hands, we lose the GFP fluorescence with DAB but we can stain for GFP and visualize GFP expression with DAB.
To do a costain, not need for DAB, simply go ahead with fluorescence staining.