Azidopine binding and pgp

[Geri Grant]

Hi,

I have been trying to get a [3H]-azidopine binding assay to work on 
membranes isolated from cell lines, and have had no success.
I get clean 170 bands on western blots using C219, but when I try to bind 
the [3H]-azidopine and run a PAGE gel, I get no signal, just a wave of 
radioactivity at the base of my gel.

Has anyone any ideas on what I may be doing wrong?

Thanks in advance.
Geraldine.