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<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>Hi
all,<o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>I
wonder if anyone has looked at the effect of Tween 80 on the s distribution. I
remember that John Philo once asked the similar question and had calculated s
for Tween 80 around 2s. <o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>I
have an 18KD protein that runs at about 1.7s when there is no Tween 80 in the
buffer. However, in the presence of Tween 80, there were two species 1.2 and
1.5s (without regularization. If F=.68, then two peaks are overlapped).<o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>The
experiment was run at 60K rpm, absorbance at 276nm. Also, the fit was not good
with rmsd above 0.01. In addition, I tried to scan at 289nm, where the
absorbance of Tween 80 is minimal, with higher protein concentration (5mg/ml).
Although the 1.2s peak is not there anymore, the main peak is at 1.4s instead
of the expected 1.7s. Also the fit was quite bad with rmsd around 0.03.<span
style='mso-spacerun:yes'> </span>For both experiments, I had 0.02% Tween in
the reference buffer.<o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>Is
it possible that the 1.2s peak is due to the Tween absorbance at the region
between the meniscuses of reference and sample sectors? I have not tried with
the same column height in both sectors. It is hard to match them exactly. Would
it be better to run this kind of experiment without including Tween 80 in
reference buffer and note the potential 2s peak as the Tween peak? For my
protein of 1.7s, it could be still difficult. Any input will be appreciated.<o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'><o:p> </o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'>Qin
"Chin" Zou<o:p></o:p></span></font></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><span
class=GramE><font size=2 face="Courier New"><span style='font-size:10.0pt;
font-family:"Courier New"'>Eli Lilly and <st1:place w:st="on">Co.</st1:place><o:p></o:p></span></font></span></p>
<p class=MsoNormal style='mso-layout-grid-align:none;text-autospace:none'><font
size=2 face="Courier New"><span style='font-size:10.0pt;font-family:"Courier New"'><o:p> </o:p></span></font></p>
<p class=MsoNormal><font size=2 face=Arial><span style='font-size:10.0pt;
font-family:Arial'><o:p> </o:p></span></font></p>
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