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MicroScale Thermophoresis – NanoTemper Technologies Monolith Instruments

MicroScale Thermophoresis

NanoTemper Technologies Monolith Instruments

MicroScale Thermophoresis (MST) measures changes of the mobility of molecules in microscopic temperature gradients. The technology is exceptionally sensitive since it detects changes in size, charge and the hydration shell of molecules. MST detects interactions between any kind of biomolecules thus providing a large application range, from ions and small molecules to high molecular weight and multi-protein complexes. Since MST is performed in a solution without any surface immobilization, bulky or sensitive molecule assemblies such as liposomes, nanodiscs or membrane proteins can be investigated.

NanoTemper provides three Monolith series instruments based on MST:

  • Monolith NT.115 and NT.115Pico: Measures thermophoresis of fluorescently labeled molecules
  • Monolith NT.LabelFree: Measures thermophoresis by intrinsic tryptophan fluorescence
  • Monolith NT.Automated: Takes up to 96 samples and combines 2 detectors in one device

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The Monolith NT.115 combines the versatility to use of a broad range of fluorophores and fluorescent proteins with all benefits of the MicroScale Thermophoresis technology. The instrument is equipped with one Nano detector that has two different fluorescence channels. The Monolith NT.115 allows measuring every interaction type – ranging from the formation of huge protein complexes to the binding of single metal ions. As there are no prerequisites with respect to the buffer composition, even such complex liquids as serum or lysates can be used for interaction studies.

Key Features of Monolith NT.115

  • Dynamic range: nM to mM dissociation constants (for sub-nM affinities see NT.115Pico)
  • Many commercially available fluorophores and fusion proteins can be used
  • Low sample consumption: just 4 µl per titration point
  • Only 10 nM of the fluorescent molecule is necessary
  • Purification Free interaction studies in lysate or serum
  • No size limitation: Monitor binding of low MW molecules as well as binding of ribosomes
  • Get your Kd in minutes
  • Kd-Plus: Get unique information on sample quality and aggregation along with the Kd
  • Maintenance-free instrument
  • Straightforward handling: Simple sample preparation and intuitive software user interface

With its high sensitivity detector the NT.115Pico is able to detect concentrations of red fluorescent dyes as low as 10 pM expanding the affinity resolution to extremely high affine interactions with dissociation constants as low as 1 pM. Since it is possible to use complex bioliquids such as lysate or serum, dissociation constants can be determined under close-to-native conditions and Purification Free.

Key Features of Monolith NT.115Pico

  • Impressive dynamic range: pM to mM dissociation constants
  • Extremely high sensitivity: Only 10 pM of the fluorescent molecule is necessary
  • Low sample consumption: Just 4 µl per titration point
  • Purification Free interaction studies in lysate or serum
  • No size limitation: Monitor binding of low MW molecules as well as binding of ribosomes
  • Get your Kd in minutes
  • Kd-Plus: Get unique information on sample quality and aggregation along with the Kd
  • Maintenance-free instrument
  • Straightforward handling: Simple sample preparation and intuitive software user interface

In contrast to the NT.115 Series instruments, the NT.LabelFree utilizes the intrinsic tryptophan fluorescence for detection allowing truly label-free and immobilization-free experiments. Without the need for a fluorescent label or fusion protein the interaction studies performed with the NT.LabelFree reflect binding of the protein in its native state. Besides the dissociation constant, MST datasets contain unique information about aggregation and sample quality allowing for rapid assay setup.

Key Features of Monolith NT.LabelFree

  • Truly label and immobilization free
  • Dynamic range: 10 nM to mM dissociation constants
  • Application Range: Ions, nucleic acids, small molecules and sugars binding to proteins
  • Low sample consumption: Just 4 µl per titration point
  • No size limitation: Monitor binding of low MW molecules as well as binding of ribosomes
  • Get your Kd in minutes
  • Kd-Plus: Get unique information on sample quality and aggregation along with the Kd
  • Maintenance-free instrument
  • Straightforward handling: Simple sample preparation and intuitive software user interface

The Monolith NT.Automated comprises all benefits of the MicroScale Thermophoresis technology and the experimental setups as applied for the Monolith series. The Monolith NT.Automated however is now able to accommodate 96 samples at a time including a robot-compatible sample preparation and reduces the overall handling time by more than 90%. The design of the Monolith NT.Automated is suited for different experimental setups; 96 samples could be used to determine 8 x 12-data point binding curves but also to perform screening assays with 96 single data points. In addition, the Monolith NT.Automated provides great versatility in the interactions to be investigated as this device can be equipped with up to two different detection systems.

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