[Histonet] Invitrogen A6455 rabbit anti-GFP dilution for immunofluorescence

[Bass, Caroline]

Hi Everyone,

I use Invitrogen/Life Technologies/Molecular Probes' antibody for GFP staining in PFA fixed rat brains. It works quite well in my hands at a concentration of 1:20,000 for DAB staining. However, I have need of immunofluorescence now and don't have a good starting point. The product sheet isn't very helpful and Invitrogen says to use the same Ab concentration which seems crazy. Almost everything I see shows a lower dilution with IF. 

Does anyone have experience with this particular antibody in the brain? If not, what's a good rule of thumb when trying to convert an antibody from DAB to IF? What would you suggest in terms of dilution?

Thanks!

Caroline