I want to determine Kd for a protein/protein interaction by ELISA. The problem is that the interaction is weak so I can not do an experiment where I let my proteins reach equillibrium in solution and thereafter meassure the unbound fraction as the proteins will dissociate directly.
I was under the assumption that I could determine Kd for a protein/protein interaction by coating one on an ELISA plate followed by titration of the other and Kd would be at the concentration corresponding to half the maximum response. Am I totaly wrong?
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