[RASMB] manual or guide of data analysis

[Cole, James]

Dear Ping

First, I'd be cautious about starting with an equilibrium measurement unless you are sure that the sample is homogeneous. Otherwise you can easily be mislead. We recommend always doing velocity measurements at several concentrations to check homogeneity and potential mass-action self-association. Sedimentation equilibrium measurements may then be useful to obtain accurate molecular weights; however, we usually find that the velocity data is sufficient and rarely go on to do equilibrium runs anymore.

You might check out some review articles to get a more detailed account of the theory practice and pitfalls of AUC:
Cole, J.L., Lary, J.W., Moody, T. and Laue, T.M. (2008) Analytical Ultracentrifugation: Sedimentation Velocity and  Sedimentation Equilibrium. Methods Cell Biol. 84, 143-179
Schuck, P. (2007) Sedimentation Velocity in the Study of Reversible Multiprotein Complexes, Protein Interactions 469–518 (a book chapter)
Laue, T. M., and Stafford, W. F. (1999) Modern applications of analytical ultracentrifugation, Annu Rev Biophys Biomol Struct 28, 75–100.

If you do decide to do equilibrium runs, we have written a software package, HeteroAnalysis, that does everything that Winnonlin/Winmatch did but is much easier to use and is more powerful. You can get it at: http://www.biotech.uconn.edu/auf/?i=aufftp

Once you get some experience you might benefit from one of our workshops. They are typically held in June but we are not doing one this year because of the AU2012 meeting workshops that were held in March. Check for updates on the RASMB website for the next one.

Good luck with your experiment,
Jim Cole

On Jun 4, 2012, at 12:22 PM, Xin-Ping Xu/FS/VCU wrote:

Hi, Everybody,
I am pretty fresh-hand in doing AUC, especially in AUC data analysis. Now I am doing Sedimentation Equilibrium, and collect some data, but I just know very little about the analysis software "Winmatch" Winnolin", is there someone who can give me some suggestions? Or whether someone has manual how to use this kind software?
Thanks,
Ping
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