[RASMB] varying temperature during an equilibrium run

[Fried, Michael G]

Hi Frank,

We tried that some time ago and found that the key was to have lots of patience for attainment of equilibrium and of course, a very stable sample.  Here's a description:

 M. A. Daugherty, M. Brenowitz,  and M. G. Fried (2000) Participation of the Amino-Terminal Domain in the Self-Association of the Full-Length Yeast TATA Binding Protein.   Biochemistry 39, 4869-4880.

Good luck!

Mike Fried

________________________________
From: rasmb-bounces at list.rasmb.org [rasmb-bounces at list.rasmb.org] on behalf of Frank Niesen [dr.frank.niesen at gmail.com]
Sent: Friday, October 25, 2013 10:47 AM
To: rasmb at rasmb.org
Subject: [RASMB] varying temperature during an equilibrium run

Hello,

I was wondering if anybody has experience with changing the temperature in an equilibrium run at given speed, to investigate temperature-dependent strength in association? Is it feasible/sane to record data at equilibrium at, e.g., 20C, and then to change the temperature to, e.g., 40C, and to hope to get data for equilibrium and record once more?

In my case, the association seems to weaken substantially with temperature and I observe a weaker curvature at the higher temperature. For a third step, going back to the original temperature, I programmed the run, but made two mistakes with it - firstly to not allow sufficient time for the equilibrium and, secondly, to somehow got the "stop AUC after last scan" checked :o(
As result, I am looking at a scan that doesn't resemble the one at lower temperature, yet it is similar to scans I saw during the equilibration between the two temperatures at step 2.

Has anybody tried something similar? Am I onto something believable/real, or shouldn't waste any more precious sample on a proper repeat (i.e. separate runs/samples for each temperature)?

Thanks in advance!
Frank
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