[RASMB] DTT for equilibrium runs?

[Leech, AP]

Debby Pheasant wrote:
> Hi, everyone,
> 
> I just heard recently that it's not good to have DTT in your buffer for 
> doing equilibrium runs.  Can someone explain to me why that would be?  
> Or share stories of DTT use?
> 
> Thanks very much,
> 
> Debby
> 
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In addition to Borries' comments I would mention that that over the
run the oxidation state of the DTT seems to "wander", so the baselines
effectively might be anywhere. Of course this may also mean that your
protein's redox state is changing. This makes the data analysis a
nightmare. TCEP is better but still seems to have some absorbance.

If you can get rid of the DTT it will make things much easier!
Ask your collaborator if it is really, really needed, sometimes
things are included in buffers just because it was grandma's recipe...

Andrew

-- 
Dr Andrew Leech                   *  Head of the Molecular
Technology Facility               *   Interactions Laboratory
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