Yes that is very true. But I assume true membrane proteins exclude high throughput :-)
However the cytoplasmic part might be fine in this case and then I would just go for Sypro Orange. Jürgen On Jul 19, 2012, at 10:39 AM, Edwin Pozharski wrote: My understanding is that the advantage of the thermofluor assay is that you can test many conditions rapidly unless of course you have some kind of high throughput CD spectrometer in mind. > If you have a CD available (not the one with music on it) you don't need a > dye just sufficient protein and you can thermal denature your protein > assuming it contains some secondary structure elements. > > Jürgen > > > On Jul 19, 2012, at 4:26 AM, anita p wrote: > > Hi All, > I want to use a thermofluor for the thermal shift assay. My proteins are > cytoplasmic truncations of membrane protein. I have read about ANS, > sypro-orange and CPM. Which is the once that is popularly used by the > crystallographers for condition optimization for crystallization ?? > > I have read that it sypro orange is not good for hydrophobic proteins and > CPM can't be used with DTT or bME in the buffer. > I am a bit confused . > Please help > thanks in advance > Anita > > ...................... > Jürgen Bosch > Johns Hopkins University > Bloomberg School of Public Health > Department of Biochemistry & Molecular Biology > Johns Hopkins Malaria Research Institute > 615 North Wolfe Street, W8708 > Baltimore, MD 21205 > Office: +1-410-614-4742 > Lab: +1-410-614-4894 > Fax: +1-410-955-2926 > http://lupo.jhsph.edu > > > > > -- Edwin Pozharski, PhD University of Maryland, Baltimore ...................... Jürgen Bosch Johns Hopkins University Bloomberg School of Public Health Department of Biochemistry & Molecular Biology Johns Hopkins Malaria Research Institute 615 North Wolfe Street, W8708 Baltimore, MD 21205 Office: +1-410-614-4742 Lab: +1-410-614-4894 Fax: +1-410-955-2926 http://lupo.jhsph.edu
