UNIT 17.8 Isothermal Titration Calorimetry

  1. Adrián Velázquez-Campoy,
  2. Hiroyasu Ohtaka,
  3. Azin Nezami,
  4. Salman Muzammil,
  5. Ernesto Freire

Published Online: 1 SEP 2004

DOI: 10.1002/0471143030.cb1708s23

Current Protocols in Cell Biology

Current Protocols in Cell Biology

How to Cite

Velázquez-Campoy, A., Ohtaka, H., Nezami, A., Muzammil, S. and Freire, E. 2004. Isothermal Titration Calorimetry. Current Protocols in Cell Biology. 23:17.8:17.8.1–17.8.24.

Author Information

  1. Johns Hopkins University, Baltimore, Maryland

Publication History

  1. Published Online: 1 SEP 2004
  2. Published Print: JUN 2004


In the last two decades, isothermal titration calorimetry (ITC) has become the preferred technique to determine the binding energetics of biological processes, including protein-ligand binding, protein-protein binding, DNA-protein binding, protein-carbohydrate binding, protein-lipid binding, and antigen-antibody binding. In this unit several protocols are presented, ranging from the basic ones that are aimed at characterizing binding of moderate affinity to advanced protocols that are aimed at determining very high or very low affinity binding processes. Also, alternate protocols for special cases (homodimeric proteins and unstable proteins) and additional information accessible by ITC (heat capacity and protonation/deprotonation processes coupled to binding) are presented.


  • isothermal titration calorimetry;
  • thermodynamics;
  • binding;
  • macromolecule-macromolecule interactions