I am currently trying to get an EMSA using TATA binding protein to work but at the moment I cannot even see a shift using a positive control!
After speaking to a couple of collegues we are thinking it could be down to the binding buffer.
Is there a basic recipie or does it need to be taylor made for different proteins.
The one that I have used consists of, 100mM Tris, 500mM NaCl, 10mM DTT with 25mM DTT/2.5% Tween-20 added seperately into the mixture.
Any help or even a standard type protocol would be great, thanks!