Immunofluorescence with FOXO - anyone done it? (Oct/01/2006 )

I've been trying to visualize the translocation of FOXO, a forkhead family transcription factor, from nucleus to cytoplasm by immunofluorescence. All I've been getting time after time is a VERY VERY dim green staining through the cell (not cyto or nuc specific). I've tried different dilutions of the antibody down to 1:50! I've also tried different dilutions of secondary (down to 1:500) Nothing . I have virtually zero experience with immunofluorescence so I'm not sure where to go from here...so I've decided to blame it on the antibody and I've ordered it from another company.

If I try to use it for a western and it works, does that mean, it should work for IF too?

Anyone else worked with FOXO?

Anyhelp would be appreciated.

I tried it as well but had also no luck...
But at least I am no IF specialist too and so I gave it up. I had so many unspecific bands in WB that I think it is an Ab problem!

Best wishes...

If I try to use it for a western and it works, does that mean, it should work for IF too?