The University of Massachusetts Amherst

Nanotemper Monolith Thermophoresis

Nanotemper Monolith thermophoresis instruments (one for intrinsic fluorescence reporters and one for extrinsic reporters)

Microscale Thermophoresis (MST) is a powerful method to quantify biomolecular interactions. It measures the motion of molecules along microscopic temperature gradients and detects changes in their hydration shell, charge or size.

In an MST experiment, a microscopic temperature gradient is induced by an infrared laser, and the directed movement of molecules is detected and quantified using either covalently attached dyes, fluorescent fusion proteins or intrinsic tryptophan fluorescence.

The applications range from small-molecule binding events to protein-protein interactions and interactions of multi-protein complexes.

Broad application range

  • access affinities (Kd, dissociation constant): pM/nM to mM range
  • examine: proteins, peptides, ribosomes, ions, nucleosomes, liposomes and more
  • study membrane proteins directly in liposomes or detergent solution
  • study multi-component reactions such as ternary complex formation, order of assembly, interfering factors, cooperativity
  • measure in any buffer, including complex detergent mixtures
  • examine interactions under physiological assay conditions such as serum, cell lysate
  • study stoichiometry and discriminate between different binding sites on a target
  • study the binding energetics ΔG (free energy), ΔH (enthalpy) and ΔS (entropy)