[Histonet] VEGF

[Amos Brooks]

Hi,
    VEGF is driving me nuts. I was hoping someone might have some
suggestions. I have been using clone VG1 for a while. I have never really
been particularly happy with it. The labeling is not nearly as specific as
I would like and there is almost always some background. If I dilute it
more the signal dies off well before the background. I've been retrieving
in Tris-EDTA for a half hour, diluting the primary at 1:30 and detecting
with Envision (Dako) and DAB.
     Recently I tried a rabbit monoclonal (I usually love these) from
Biocare Medical. I can certainly say it isn't as dirty as the VG1 clone,
unfortunately there is not much signal at all. The recommended dilution is
1:100 to 1:200. I have tried as low as 1:50. Biocare tech support (nice
guys) suggested retrieving for an hour which I tried. I have been
increasing the incubation time from a half hour to 90 min to overnight in
the fridge.
     Honestly guys, there's not much hair left to pull out. Does anyone
have any suggestions? I'd really like to get this working properly.

Thanks,
Amos