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How does the specificity to bind to antibody differs between 'hot' and 'cold' antigen in radioimmunoassay? This question is in relation to radioimmunoassay. In this assay, the 'cold' antigen replaces the 'hot' antigen. Why is it so, if both the antigens are same and hence their specificity? Does the radioactive molecule change its specificity?

The specificity is the same, but protein-ligand binding is an equilibrium process. In other words, the antigen is constantly associating with and dissociating from the antibody. As a hot antigen dissociates, it is possible that a cold antigen can take its place, and vice-versa. As the ratio of cold to hot antigen increases, there is a greater probability that cold antigens will take the place of hot antigens.

You can read more on Wikipedia.

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