Dear Theresa, Although I haven't yet done the experiment myself, I am told that microscale thermophoresis (MST) is another useful method for oligomerization measurements, especially since it is designed to work for membrane proteins in presence of detergents etc. If you want to quickly learn more about microscale thermophoresis, you can google the technique for further information and literature. If you end up wanting to try MST experiments and don't have much of a handle on the method yet, contact me off the bulletin board and I'd be happy to give you a few pointers.
As far as Thermofluor goes, some of the detergents work better than others in Thermofluor experiments so you may have it to determine the issues in your case empirically. Good luck! Raji On Thu, Mar 21, 2013 at 2:10 PM, Theresa Hsu <[email protected]> wrote: > Dear all > > I have a His-tagged membrane protein with unknown oligomerization state. > But I am worried that tag addition may induce different state than in > native and affect its crystallizability. > > Is there a single method that can determine the oligomerization state with > nearly 100% accuracy? I have use of AUC and SAXS but there seems to be > ambiguity about detergent and lipid effects. Is Thermofluor a right method? > > Does oligomerization require special assembly proteins, which will mean > that tag cleavage is not useful to obtain native state? > > Thank you. > > Theresa > > -- Raji Edayathumangalam Instructor in Neurology, Harvard Medical School Research Associate, Brigham and Women's Hospital Visiting Research Scholar, Brandeis University
