MST - Microscale Thermophoresis
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MST in in Biochemistry, Biophysics and Structural Biology
Microscale Thermophoresis (MST) is a biophysical technique used to study molecular interactions by detecting changes in the movement of biomolecules and complexes along a temperature gradient.Ìý
When subjected to a localised temperature increase, molecules experience a shift in their mobility, a process known as thermophoresis. The extent of this movement is dependent on the size, charge, and hydration shell of the molecule or complex. During most binding events, one or several of these properties will change, making MST a powerful technique for the quantitative analysis of binding reactions.Ìý
By labelling one of the interacting partners with a fluorescent dye, MST monitors changes in fluorescence intensity as the labelled molecule moves in response to a small temperature gradient. The highly localised temperature gradient is induced with an infrared laser (1480 nm wavelength) focused onto a small spot on the sample capillary where the fluorescence intensity is measured. When binding occurs, the thermophoretic properties of the labelled molecule change, enabling the detection and quantification of molecular interactions based on changes in fluorescence intensity due to thermophoretic movement. By analysing these shifts over a series of different concentrations, MST provides detailed information on binding affinities.
MST can measure dissociation constants (Kd) from the nanomolar (nM) to millimolar (mM) range. It is applicable to various interactions, including protein-protein, protein-ligand, DNA/RNA-ligand, DNA/RNA-protein, protein-lipid, peptide-lipid, small molecule binding, and membrane proteins in liposomes, nanodiscs, or detergent solutions.
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Key highlights of MST in Biology, Biochemistry, and Biophysics
- Dissociation constants (Kd) ranging from nM to mM.
- Measurements under equilibrium conditions in solution.
- Very few buffer restrictions; measurements possible in complex buffers or even in lysates under near-physiological conditions.
- Applicable to various interactions: protein-protein, protein-ligand, DNA/RNA-ligand, DNA/RNA-protein, protein-lipid, peptide-lipid, small molecule binding, and membrane proteins in liposomes, nanodiscs, or detergent solutions.
- No immobilisation and no size limitations.
- Flexible labelling options, with the label positioned far from the interaction site to minimise interference with binding.
- Low sample consumption.
- Determine stoichiometry and number of binding sites.
- Investigate oligomerisation and aggregates.
Instrument and Accesories
Our MST instrument is a Nano-BLUE/RED Monolith NT.115 from NanoTemper Technologies.Ìý
It has two LED-filter combinations:
- blue (excitation 450–480 nm, emission 515–570 nm) Ìý
- red (excitation 600–645 nm, emission 670–730 nm)