[RASMB] 2 subjects, overspeed, " SEGAL"
Lustig
ariel.lustig at unibas.ch
Wed Jun 26 08:50:00 PDT 2002
Dear RASMB colleagues ,
Dear Andrew,
Subject 1
in the "Manual of Methodes" (C H Chervenka) for the AUC Spinco -Beckman
Division-Palo Alto
(for the model "E") is the Overspeed -technic discussed, page 44 and ref.75:
Hexner P E, Radford L E ,Beams J W "Achievement of SE "
Proc. Natl. Acad. Sci. 47, 1848 (1961)
As you ask for a practical procedure=rule of thumb, I like first of all to
emphasize
that the time to reach equilibrium depence on three parameters, 1)The
filling height
of the cell, 2) the diffusion constant of the solute=substance . 3) the
viscosity of the solution.
The optimal over-speeded rotor speed and time is, that still at least a
1/3 to 2/3 of the plato,
(the region that in case of absorption shows approximately the
C(0)=initial concentration
you filled and tested it at very low speed , just at the run -start or
compaire it with the wavelength
scan at the same nM value.(Be a bit carefully about the WL baseline.)
What are the dangerous points on have to be careful : at a too relative
high speed the sample
may be dragged too far to the cell bottom , so you loose the time
-spare advantage,
but worth is if the particles
or molecules du not reversibly back-diffuse (means insoluble aggregates)
due to high
concentration near the bottom.
On principle we have to look several times during the overspeed -
procedure on the partly
depleted part near the meniscus and the unreached bottom cell part.
Subject 2.
I would like to remind you that at the 23 of may 2002 I sent to all of
you our =(Biozentrum)
SEGAL computer that can be loaded down from the net
http://www.biozentrum.unibas.ch/personal/jseelig/AUC/index.html
I would like to hear from you if there are some difficulties loading it
down or to use it.
please inform me .
WE can do a play:
You send me a single SE(at equilibrium ) scan by E-mail of one of your
scans with
the information of V = the part.spec. volume and the solution density.
BUT PLEASE
DON'T give me the Mw. Later we will compaire with your result from the
same scan.
Thanks to all of you ....ariel
Lustig Ariel
Dept. of Biophysical Chemistry / Analytical Ultracentrifuge Lab.TEL....+61
2672187
Biozentrum der Universität
Basel Fax +61 2672189
email
lustig at ubaclu.unibas.ch
Klingelbergstr. 70
CH 4056 Basel
Switzerland
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