I have a similar case, where in there are multiple binding sites on the protein for the ligand and ligand induces dimerization.So it is not helpful even if I separate the monomer and dimer. If I titrate the dimer with ligand, the stoichiometry will completely change? Any suggestions will be helpful.
On Fri, Jul 18, 2014 at 12:46 PM, David Briggs <[email protected]> wrote: > Hi Sajid, > > *Assuming* you have one site per monomer (rather than, say, one site per > dimer), and *assuming* each binding event is completely independent ( I.e > no co-operativity), you might just get away with running the experiment > with the heterogeneous material. > > However, you might not be able to confidently make these assumptions, so > imho it would be preferable to separate the monomer and dimer by SEC prior > to ITC. If this is not possible, then pay close attention to the fit when > you run the heterogeneous experiment. Poor fit to a one site model may > indicate that these assumptions are invalid. Can you obtain stoichiometry > information from a different technique? This might be very helpful. > > Hth, > > Dave > > Dr David C Briggs PhD > http://about.me/david_briggs > On 18 Jul 2014 10:25, "sajid akthar" <[email protected]> wrote: > >> Dear All, >> >> This is an off-topic question. I have protein solution of heterogeneous >> (contains both monomer and dimer). I want to perform ITC with this protein. >> I doubt whether this heterogeneity will interfere the binding study. >> >> Any advice please. >> >> Thank you >> >> Sajid >> >
