Nano Isothermal Titration Calorimeter from TA instrument (Karditsa)
Isothermal titration calorimetry (ITC) can determine all the thermodynamic parameters (affinity, enthalpy and stoichiometry) of a binding interaction (e.g. A + B ↔AB) in one experiment. 1. ITC works by titrating reactant B into reactant A under isothermal conditions.
User should supply a sample usually purified protein (reactant A) and a ligand (reactant B)
Requirement
Reactant A
Purity > 90%
Concentration: ideally 30 * Kd (e.g. for a Kd around 0.5 μΜ sample concentration should be 15μM around)
Usually working with concentration between 10-30μM
Volume: 1.2ml (ideally for a sample and blank experiments 2.5 ml)
Reactant B
Concentration: 15 to 20 * n * C where n is the stoichiometry of the reaction (A + B ↔AB) where C concentration of reactant A
Volume: 0.1ml (ideally for a sample and blank experiments 0.2 ml)
DESCRIPTION:
FUNDER: