[Histonet] DAB GFP staining

[koellingr <@t> comcast.net]

Alonso,

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 -------------- 
From: alonso.martinezcanabal <@t> utoronto.ca 

> 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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