Isothermal Titration Calorimetry – Advantages

Isothermal Titration Calorimetry – Advantages

In general, ITC is the go-to method for determining high-quality thermodynamic parameters of molecular interactions. Moreover, it provides a direct, label-free, and in-solution approach.

No label required
→ close to native molecular state/conformation

 

No immobilization required
→ No influence on molecular conformation/stability

 

Wide temperature range
→ 10 to 80 °C

 

Wide range of possible molecules and ligands
→ No limitation in molecular weight

 

Wide range of possible buffers and solvents
→ Only few additives may cause background heat signals

 

Relatively low sample consumption for ITC standards
→ Only 280 µL of macromolecule (at around 20 µM)
→ Only 40 µL of ligand (at around 200 µM)

 

Complete thermodynamic characterization in one experiment
→ Enthalpy, entropy, and free enthalpy of binding

 

Determination of binding characteristics
→ Dissociations constant and reaction stoichiometry

 

Wide range of binding affinity measurable
→ Millimolar to nanomolar dissociation constants