Sample guidelines beta

These guidelines are for orientation only, please check carefully before your experiment.

An example of a preliminary experiment is to inject aliquots of the titrant (40 µL at concentration 10 times higher than the protein) into the sample cell containing the protein solution (220 µL, at about 10-100 µM).

A real ITC experiment consists of two titrations, one where the titrant (syringe) is injected to the macromolecule (sample cell); and a second one, where the titrant is injected to the buffer. Please check below for other examples of experiments.

scientist using a pipette in a lab
Ultra-sensitive iTC200 with reduced ligand and sample cell volumes.
Experiment/Parameter Affinity Kd Protein concentration Ligand concentration
Experiment 1 200 µM 75 µM 7 mM
Experiment 2 2-20 µM 30-60 µM 0.5-1 mM
Experiment 3 2 nM 25 µM 0.15 mM
Table 1: Examples of experiments performed at the iTC200 microcalorimeter.
Use these values only as a guide, sample concentrations might need to be optimised for each case study.

Analysis of interactions by calorimetry (ITC)

ITC is the most precise technique to measure the affinity between macromolecules such as proteins, nucleic acids, or any small molecule.