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Calcineurin is a Ca2+/calmodulin-dependent phosphatase. It is very important to study the affinity between calcineurin and its substrate or other interacting proteins. There are two conserved motifs on calcineurin-interacting proteins, namely PxIxIT motif and LxVP motif. Microscale Thermophoresis (MST) is fast and accurate in determining the affinity between protein-peptide interactions.