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<DIV dir=ltr align=left><SPAN class=370283215-25102013><FONT color=#0000ff
size=2 face=Arial>Frank, I haven't actually done that type of protocol, but in
principle it should work (assuming the protein is stable enough). My comment
though is that if the association is weaker at higher temperatures you want to
do the higher temperature <U>first</U> and then the lower (for the same reason
you always want to go up in speed for a multi-speed SE sequence). It takes much
longer to reach equilibrium when the change (temperature or speed) will
produce a gradient that is less steep than when the gradient
increases.</FONT></SPAN></DIV>
<DIV dir=ltr align=left><SPAN class=370283215-25102013><FONT color=#0000ff
size=2 face=Arial></FONT></SPAN> </DIV>
<DIV dir=ltr align=left><SPAN class=370283215-25102013><FONT color=#0000ff
size=2 face=Arial>John</FONT></SPAN></DIV>
<DIV lang=en-us class=OutlookMessageHeader dir=ltr align=left>
<HR tabIndex=-1>
<FONT size=2 face=Tahoma><B>From:</B> rasmb-bounces@list.rasmb.org
[mailto:rasmb-bounces@list.rasmb.org] <B>On Behalf Of </B>Frank
Niesen<BR><B>Sent:</B> Friday, October 25, 2013 7:47 AM<BR><B>To:</B>
rasmb@rasmb.org<BR><B>Subject:</B> [RASMB] varying temperature during an
equilibrium run<BR></FONT><BR></DIV>
<DIV></DIV>
<DIV dir=ltr>Hello,
<DIV><BR></DIV>
<DIV>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?</DIV>
<DIV><BR></DIV>
<DIV>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(</DIV>
<DIV>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.</DIV>
<DIV><BR></DIV>
<DIV>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)?</DIV>
<DIV><BR></DIV>
<DIV>Thanks in advance!</DIV>
<DIV>Frank</DIV></DIV></BODY></HTML>