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J.Dunn 07-29-2013 08:37 AM

Binding buffer
 
Hi
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!


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