In article <[email protected]>, [email protected] says... > > Hello folks > > I would like to do protein conformation stability test, which I can do with > normal chemicals, which is available in any common lab. Some where I have > found that protein+urea+DTT incubated at room temperature and load on > SDSPAGE/Native page. But I have not understood very well of the protocol. > > If anyone has information please let me know > > Thank you in advance
You cannot measure protein unfolding by SDS-PAGE, as SDS itself unfolds proteins. Native PAGE may work. Indeed, you could use a urea gradient across a gel to measure unfolding (Creighton 1979). SDS-PAGE followed by Western are used to measure unfolding by pulse proteolysis (Kim et al., 2009). Any other suitable method to measure protein structure may also be used to generate "chevron plots" (Fersht & Matouschek 1991, Zarrine-Afsar & Davidson 2004). If you have enough material, differential scanning calorimetry may give the best results. _______________________________________________ Methods mailing list [email protected] http://www.bio.net/biomail/listinfo/methods
