Hi Anita, You don't state the WL of the melting and "back folding" CDs, I'm guessing it's around 210 nm?
In any case, the "refolding" that you see might be just the formation of aggregates. For saying if it's really refolding you'd need a new spectrum WL vs AU in the end of the "reverse thermal melt" and superimpose it with the first one. Having the protein more concentrated than that, or using a longer path length could reduce your noise. Carlos Em 14/05/2012, às 09:10, anita p escreveu: > Hi All, > Sorry for this off topic. > I am stuck with a plant protein which is not crystallizing. I am expressing > this in E coli. > I have done the CD and thermal melt of the deletion constructs of this > protein @ 0.2 mg/ml. > > Please refer to the doc file attached to this email. > > The CD looks okay but the thermal melt looks kind of wierd. I would expect a > sigmoid curve for the thermal melts and then when we do a reverse temperature > scan, the protein should denature and may not fold back (with some exceptions) > > Please let me know if any one had the same experience before and could > explain me what this meant. > > Kind regards > Anita > <Cd.doc>
