Plan Your Project

MST experiments allow us to measure binding affinities between molecules, such as protein-ligand interactions, by detecting changes in the mobility rate of molecules in response to a thermal gradient.

MicroScale Thermophoresis method, is used to detect changes in the hydration shell, charge or size of molecules. The instrument uses an infrared-laser to generate a small temperature gradient within thin glass capillaries that are filled with the sample. The motion of molecules along the temperature gradient is monitored by the fluorescence of the molecules. The change of mobility rate along the temperature gradient can be used to determine binding affinities, binding kinetics, and activity kinetics.

We can supply with the following protein labeling kits by NanoTemper, which are optimized for MST: His-tag, NHS (amine), or cysteine labeling. You can use this 'Labeling Decision Tree', by NanoTemper, to help determine the best labeling strategy for your experiment.

Yes. Our instrument, the Monolith NT.115, is equipped with two different fluorescence channels, BLUE/RED. Any fluorophores that can be detected within this range, such as GFP and Cy5, can be suitable for our instrument. Here, you can find a list of the compatible fluorophores.

Each MST measurement takes no longer than 15 minutes. The overall time, including labeling and sample preparation, typically ranges from 1 to 2 hours, depending on the complexity of the interaction and the number of measurements required.

The concentration of the labeled molecule typically ranges from 10 to 100 nM. If you are using NanoTemper labeling kits, the required volume and concentration will be determined based on the chosen labeling strategy.

The concentration of the unlabeled molecule depends on its affinity. If the expected Kd is unknown, we typically start with a concentration of 1 mM. Based on the results of the first experiment, we can adjust the concentration to obtain a sigmoid curve.

Note: If the concentration of the labeled molecule is lower than the Kd, its precise concentration is not critical.

For each binding experiment you will need 180ul of the labeled molecule, 30ul of the

unlabeled molecule and 180ul of its buffer. It is important to bring higher volumes of each component for the initial tests.