Directly measure the strength of molecular interactions in solution, without the need for immobilization. Characterize nearly all types of molecular interactions with ultra-low sample consumption and versatile capabilities
Monolith X uses Spectral Shift technology to quantify molecular interactions between a target and a ligand.
When the target is labeled with a fluorophore, it gives a particular emission spectrum. If a ligand binds, the fluorophore’s chemical environment is changed, causing a blue-shift or red- shift or broadening of the emission peak
Fluorescence is recorded and a 670nm/650nm ratio is calculated and plotted against the ligand concentration (bottom). This ratiometric measurement is used to derive the affinity constant (Kd) which is automatically determined at the end of each run without additional and lengthy data analysis
Signal measured at two distinctive wavelengths
Ratio of signals plotted against ligand concentration
To quantify molecular interactions with MST, start by labeling your target molecule with a fluorescent dye and mix it with your ligand. A very precise and brief laser-induced temperature change is applied, which amplifies the variation in fluorescence intensity caused by the ligand binding to your target.
The Kd is calculated by plotting the change in fluorescence against the ligand concentration
