Re: [Histonet] DAB GFP staining


What kind of sections are you using?  Are they thick enough?  Thin sections would show nuclei and not "branches" since "branches" would randomly be out of the plane of a too thin section.  Mainly used flourescent tagging of GFP cells in mouse brain with anti-GFP and a flourescent secondary.  But certainly have done this nicely with appropriate secondary and then DAB.  Used a goat anti-GFP,  (you can use others), a biotinylated horse anti-goat from Vector (I personally like Vector secondaries but can use others), avidin/Peroxidase and DAB.  You can actually see a beautiful picture on AbReviews of mouse brain stained with GFP and DAB that I have to admit outdoes anything I did yet they have a very similar procedure although I certainly didn't write the review or produce the slide but mine were pretty darn good.

Ray Koelling
PhenoPath Labs
Seattle, WA

-------------- Original message -------------- 

> Dear everyone. 
> Have someone ever tried to perform a DAB 
> immunohistochesmitry in tissue transfected with GFP? Some people in my 
> lab have been trying it with not very good results. This is 
> hippocampus and apparently just the nuclei and not the branches are 
> stained. 
> Thanks. 
> Alonso 
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