Monolith NT.115

Micro-Scale Thermophoresis (MST)

This method is used to determine the binding affinity between molecules, by measuring their directed movement along a temperature gradient (an effect termed “thermophoresis”). The Monolith systems measure equilibrium binding constants between a variety of molecules, with almost no restriction to molecular mass. It allows fast detection of a wide range of biomolecular interactions. Measurements are performed immobilization-free and in any buffer or complex bioliquid (e.g. serum and cell lysate).

Application

• Determining dissociation constant

• Determining EC₅₀

Requirements

• One of the binding partners needs to be fluorescently labeled.

  • This can be achieved either by expressing a recombinant protein fused to a fluorescent label, or by fluorescent protein labeling kits (such as: https://nanotempertech.com/monolith/consumables/#protein-labeling-kits).

• The assay is performed in specialized capillaries manufactured by NanoTemper (https://nanotempertech.com/monolith/consumables/#capillaries).

Advantages

• Immobilization-free

• Any buffer (even cell lysate or serum)

• Low sample consumption

• High dynamic range (K_d values of 1 nM to 500 mM)

• Fast

Type of interactions

• Protein-Protein

• Protein-Small Molecule

• Protein-Ion

• Protein-Vesicle

• Protein-DNA

• DNA-DNA

• DNA-RNA