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Energetics-based target identification by pulse proteolysis. A cell lysate is incubated with a ligand and urea. A control reaction is prepared in the identical way except without the ligand. After incubation, unfolded proteins are digested by pulse proteolysis and remaining proteins are analyzed by 2-D gel electrophoresis. Proteins in the spots showing differential intensities on the 2-D gels of the two reactions are identified and characterized further.
