Hello, I am young researcher working in computational drug discovery and willing to perform experimental work, for which I am planning to start with ITC. Currently, I am reading relevant literatures to develop an idea about the task but what I have been unable to find out, is the protocol for preparing the assay buffers for the protein and ligand. Can anyone refer me a good protocol?
An important detail is that the protein and ligand should be in matching buffers, so that there will be minimal heat of dilution due to differences in conditions. This can be accomplished by dialyzing the protein extensively against the experimental buffer, and then dissolving the ligand in the same dialysis buffer. The same buffer should also be used to make any dilutions.
Typically, the ligands to be titrated in ITC should be dialyzed extensively against the same buffer to minimize the heat effects of buffer mixing.
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