[RASMB] varying temperature during an equilibrium run

[Kirk C Aune]

On 10/25/13 10:47, Frank Niesen wrote:
> 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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I refer you to Dimerization of Alpha-Chymotrypsin. II.  Ionic Strength 
and Temperature Dependence.  Biochemistry 10, no.9, pp 1617-1622 
(1971).  Kirk C Aune, Lowell C. Goldsmith and Serge N. Timasheff.

This, of course, was with a Model E  AUC.  In order to get stable 
temperature control above 30C with no oil condensation, we employed an 
aluminum flashing liner on the chamber wall.

Cheers.

-- 
Kirk C Aune, Ph.D.
16289 Cambridge Court
Granger, IN 46530
(574)277-8589
Linux User #1720

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