10/2014 Monolith NT.115 Series Product Information Monolith Instruments for MicroScale Thermophoresis www.nanotemper-technologies.com 10/2014 MicroScale Thermophoresis A technology by NanoTemper MicroScale Thermophoresis is an easy, fast and precise way to quantify biomolecular interactions. It measures the motion of molecules along microscopic temperature gradients and detects changes in their hydration shell, charge or size. ▶ Judge and improve sample quality immediately ▶ Work with very small amounts and sensitive samples ▶ Analyze in all buffers and bioliquids (cell lysate, serum) –immobilization-free ▶ ▶ 2 and get rid of maintenance downtime Kd s for all molecular weights from ions to ribosomes www.nanotemper-technologies.com 10/2014 MicroScale Thermophoresis A powerful technique MicroScale Thermophoresis (MST) is a powerful technique for quantifying is induced by an infrared laser, and the directed movement of molecules is Monolith NT.115 Series | Product Information 3 10/2014 Discover the Application Range MicroScale Thermophoresis detects interactions between any kind of biomolecules thus providing a large application range, from ions and small Thermophoresis, the movement of molecules in temperature gradients, is not only dependent on the size, but also on the charge and the hydration shell of the molecule of interest. Therefore, binding events can be detected nm 0. 1 nm 7 1 nm nm nm 75 Size 20 0 10 00 nm Since MST is performed free in solution without any surface immobilization, also bulky or sensitive molecule assemblies such as liposomes, nanodiscs or membrane proteins can be investigated. MicroScale Thermophoresis (MST) Ribosomes Bacteria Cell Nucleic acids Ions Nucleosomes Nucleotides Viruses Proteins Atoms Fragments Liposomes NanoDiscs Antibodies AntibodyFragments Peptides Small Molecules 4 www.nanotemper-technologies.com Da 1 Da 15 0 1 kD a a kD 10 0 10 00 kD a MW 10/2014 Monolith NT.115 and NT.115 Pico The NT.115 Series measures biomolecular interactions via detection of ▶ ▶ ▶ ▶ Broad application range Cat # 1 2 Instrument Channel 1 Channel 2 Kd) Sample Consumption (per Kd) NT.115Pico Pico – RED - 1 pM to mM 1 NT.115 Nano – BLUE Nano – 1 nM to mM 2 NT.115 Blue/Red Nano – BLUE Nano – RED 1 nM to mM 2 NT.115 Nano – Nano – RED 1 nM to mM 2 Kd Kd Monolith NT.115 Series | Product Information 5 10/2014 Technical Details Monolith Instruments NT.115Series NT.115 NT.115Pico 2 1 Samples per run (RED) Labeling required 1 nM to mM 1 pM to mM Yes Yes -3 M -11 -3 M Range of accessible interactions Biophysical parameters Enthalpy, Enzyme Kinetics Enthalpy, Enzyme Kinetics No No Yes Yes (serum, cell lysate) Volume per measurement Molecular weight range (Da) 1 1 Minutes Minutes Immobilization required No No Temperature controlled 22 - 45 °C 22 - 45 °C Maintenance required No No 77 7 7 7 10/2014 mated NT.Automated ghput ST MST Monolith Monolith Monolith Monolith Detectors Detectors Monolith Detectors Detectors Detectors The The Monolith Monolith The Monolith Theinstruments instruments Monolith The instruments Monolith instruments harbor harbor instruments harbor different different harbor different harbor different different detectors detectors detectors to detectors to monitor monitor to detectors monitor to the the monitor fluorescence fluorescence the to monitor fluorescence the fluorescence the signal signal fluorescence signalsignal signal of of your your of sample. your sample. of sample. your The of The sample. your excitation/emission excitation/emission Thesample. excitation/emission The excitation/emission The excitation/emission wavewavewavewave-wavelengths lengths lengths for forlengths the the forcorresponding corresponding lengths the forcorresponding thefor corresponding thedetectors detectors corresponding detectors are detectors are are detectors are are illustrated illustrated illustrated below. illustrated below. below. illustrated The Thebelow. Monolith Monolith The below. Monolith TheNT.Automated NT.Automated Monolith The NT.Automated Monolith NT.Automated NT.Automated can can be be can equipped equipped be can equipped be can equipped with with beaa with equipped combination combination awith combination awith combination of of a two combination two of two of twoof two NT.115 Series detectors are illustrated below. different different different detectors. detectors. different detectors. different detectors. detectors. Detectors and Spectra seat a time grate deility: integrate 2 de2 decombine nsitivity high sensitivity high sensitivity abel-free ce e and label-free ns Label Label [nm] Free Free [nm] [nm] [nm] [nm] 800 800 800 800 800 750 750 750 750 750 700 700 700 700 700 650 650 650 650 650 600 600 600 600 600 550 550 550 550 550 500 500 500 500 500 450 450 450 450 450 400 400 400 400 400 350 350 350 350 350 300 300 300 300 300 250 250 250 250 250 Nano Nano Nano Label Nano Nano Label BLUE/ BLUE/ BLUE/ Free BLUE/ BLUE/ Free RED GREEN GREEN GREEN RED Pico Nano Nano Nano Pico Pico Nano Nano Nano Nano Pico Nano Nano Pico Label Red Red BLUE/ GREEN/ GREEN/ Red GREEN/ Red BLUE/ GREEN/BLUE/ BLUE/ Red BLUE/ GREEN/ Free RED RED RED RED GREEN RED GREEN RED RED dling id-handling into liquid-handling ated s son tectors to of 2 detectors up to 2 detectors ameters depend on the depend on the on the uhe uctors: heNT.Audetectors: The NT.AuThe NT.Au- a d quipped an with beaequipped with a with a o on ntdifferent of de-two different dede- en be yn ch chosen freely can be chosen freely chosen ms onolith netection s systems detection systems systems abelFree eodeand LabelFree dede- The Thewhite white Theand and white The black black and white shaded shaded black The and white shaded boxes black boxes and shaded correspond correspond boxes black correspond shaded boxes to tocorrespond the the boxes to the correspond to the to the excitation excitation excitation and andexcitation emission emission and excitation emission wavelength wavelength and emission wavelength andspectra spectra emission wavelength spectra of ofwavelength the thespectra of the spectra of the of the different different different fluorescence fluorescence different fluorescence different detectors, detectors, fluorescence detectors, fluorescence respectively. respectively. detectors, respectively. detectors, respectively. respectively. d) ly controled) detectors. ce cic ormance diagnostic and performance diagnostic diagnostic Monolith Monolith Monolith Monolith Instruments Instruments Monolith Instruments Instruments Instruments 10/2014 10/2014 Detection of picomolar Kd NT.115Pico. Material was kindly provided by Dr. Ute Curth, Medical Unitersity Hanover, and Dr. Ahmed Besheer, Novartis, Basel. Jerabek-Willemsen, M., André, T., Wanner, R., Roth, H. M., Duhr, S., Baaske, P., and Breitsprecher, D. 10/2014 Customer Statements Basel, Switzerland “We have used our Nanotemper NT.115 MicroScale Thermophoresis (MST) we have been able to make comparisons, results from MST agree well with anisotropy, SPR). However, we particularly appreciate two distinguishing molecules binding to large labeled molecules, where binding is tight (nM Kd) and one or both partners aggregate at micromolar concentrations. Secondly, titrations, particularly for systems that have changes in their conformational or oligomeric state upon binding. In several cases, multi-phasic titrations Prof. Dr. Uffe Holmskov, Institute for Molecular Medicine, University of Southern Denmark, Odense, Denmark thus, we appreciate that the interactions are investigted free in solution and can be studied even in cell lysate. We are using MST for a number of different interactions, mainly to investigate protein-protein interactions but and biologics projects, with NanoTemper Technologies’ MicroScale Thermophoresis has proved a valuable tool for characterising small molecule-protein and protein-protein interactions, as well as for the study of protein aggregation concentration determination. There is very good agreement with other technologies such as Surface Plasmon Resonance (SPR) and Isothermal Titration Calorimetry (ITC), and we are particularly 10/2014 Selected Publications 1. 2. 3. Jerabek-Willemsen, M., André, T., Wanner, R., Roth, H. M., Duhr, S., Baaske, P., and Breitsprecher, Structure 4. thermophoresis provides insights into mechanism and thermodynamics of ribozyme catalysis. RNA 5. Phosphoconformational Switch that Reduces Microtubule Association. Current Biology Taft, M. H., Behrmann, E., Munske-Weidemann, L.-C., Thiel, C., Raunser, S., and Manstein, D. triggered activities of the RNA-binding protein HuR. Cellular Signalling hepatitis B virus core protein. Proceedings of the National Academy of Sciences 11. Xiong, X., Coombs, P. J., Martin, S. R., Liu, J., Xiao, H., McCauley, J. W., Locher, K., Walker, P. A., 12. 1524 13. Monolith NT.115 Series | Product Information 11 10/2014 Europe / International info@nanotemper-technologies.com USA / Canada NanoTemper Technologies, Inc. info@nanotemper-technologies.com www.nanotemper-technologies.com