On Exchange Reaction
To initiate an H/D exchange reaction, a protein sample, initially in non-deuterated buffer, is incubated in a buffer with 50-90% mole fraction deuterated water. There are almost no restrictions on reaction conditions which allow exchange behavior to be studied as a function of protein and buffer composition, solution pH, and in the presence and absence of ligands. To follow the deuterium buildup of individual amide hydrogen or sets of hydrogens, several on exchange time points are sampled for each condition.
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