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Tina et al.:<br><br>
have you ever run the protein through a size exclusion column? If
you have good recovery from sizing column, I would suggest using size
exclusion in combination with light scattering, refractometry and
absorbance as no density matching is required.<br><br>
Yutaro Hayashi, Hideo Matsui and Toshio Takagi (1989) Methods
Enzymol,172:514-28 <br><br>
Yernool D, Boudker O., Folta-Stogniew E., and Gouaux E. (2003) Trimeric
subunit stoichiometry of the glutamate transporters from Bacillus
caldotenax and Bacillus stearothermophilus. <i>Biochemistry
</i><b>42</b>: 12981-12988<br><br>
Albright R A, Ibar J. L. V., Kim C. U., Gruner S. M., and Morais-Cabral
J. H. (2006) The RCK Domain of the KtrAB K+ Transporter: Multiple
Conformations of an Octameric Ring. <i>Cell </i><b>126</b>:
1147-1159<br><br>
Ewa<br><br>
At 10:50 AM 1/6/2010, Tina Daviter wrote:<br>
<blockquote type=cite class=cite cite="">Dear All,<br><br>
I will have to run an SV experiment in the presence of the detergent
ASB-14 at 0.5%. It is a solid and I wondered if anyone has any
information on the density of it in solution so I can do density
matching? Or is it not that easy in this case?<br><br>
The aim of the experiment is to get the oligomeric state of a membrane
protein.<br><br>
If the density is unavailable, any suggestion on how to most easily
approach the problem would be highly welcome. I fear to determine the
density by running ASB-14 in H20/D20 mixes might fail as it does not seem
to absorb much at any wavelength (is interference sensitive
enough?).<br><br>
Also, I have not entirely understood how to account for the influence
detergents have on the viscosity. I should be able to organise access to
an Anton-Paar densitometer, but I don't think there is a viscosimeter
that I could use.<br><br>
Is SE inavoidable?<br><br>
For more information on the detergent, if that helps, here is the full
name and link to Sigma:<br>
Amidosulfobetaine-14,3-[N,N-Dimethyl(3-myristoylaminopropyl)ammonio]propanesulfonate.<br>
<a href="http://www.sigmaaldrich.com/catalog/ProductDetail.do?D7=0&N5=SEARCH_CONCAT_PNO|BRAND_KEY&N4=A1346|SIGMA&N25=0&QS=ON&F=SPEC" eudora="autourl">
http://www.sigmaaldrich.com/catalog/ProductDetail.do?D7=0&N5=SEARCH_CONCAT_PNO|BRAND_KEY&N4=A1346|SIGMA&N25=0&QS=ON&F=SPEC</a>
<br><br>
Literature recommendations (preferably overviews and experimental
methods) are also welcome.<br><br>
Thank you very much in advance.<br><br>
Regards<br>
Tina<br><br>
<br>
-- <br>
Dr. Tina Daviter<br>
ISMB Biophysics Centre<br>
Birkbeck, Univ. of London<br>
London, UK<br><br>
T 020 70790749<br>
E t.daviter@mail.cryst.bbk.ac.uk<br>
W
<a href="http://biophysics.ismb.lon.ac.uk/" eudora="autourl">
http://biophysics.ismb.lon.ac.uk</a><br>
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