Isothermal titration calorimetry: VP-ITC
Technology / Methodology:
Interactions and characterization of biomolecules
Location:
CEITEC MU, building A4, room 2.18
Research group:
CF: Core Facility Biomolecular Interaction and Crystallization
VP-ITC is isothermal calorimeter with 1400 µl volume of working cell. ITC method is used for characterization of biomolecular interactions of small molecules, proteins, antibodies, nucleic acids, lipids and others. Enzyme kinetics, biological activity or the effect of molecular structure changes on binding mechanism can be also assessed. Complete thermodynamic profile of the molecular interaction in a single experiment (stoichiometry, Ka, ∆H and ∆S values) or kinetics parameters Km and kcat can be determined.
VP-ITC is designed to measure the heat of binding. In a typical arrangement, the titrant, also referred as the ligand, is injected into the sample cell containing the macromolecule sample material. The calorimetric measurement can be done over a range of biologically relevant conditions (temperature, salt, pH, etc.). ITC system directly measure submilimolar to nanomolar binding constants (Ka=10^3 - 10^9 M-1). Measure of the interactions with nanomolar to picomolar binding constants (Ka=10^9-10^12 M-1) can be measured using the competitive binding technique, the same principle can be used for low affinity interactions (10^3-10^2 M-1).
The operating temperature range is of 2°C to 80°C.
Measurement at VP-ITC is supposed to be performed manually by user itself after training.
For more details, please contact responsible person:
Monika Kubíčková
e-mail: monika.kubickova@ceitec.cz
phone: +420 549 49 25 27
office: 3.39/A4
