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Nano Temper Monolith NT.115 User Manual

Microscale thermophoresis instrument
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Summary of Contents for Nano Temper Monolith NT.115

  • Page 1 User Manual ® Monolith NT.115...

  • Page 2: Table Of Contents

    Table of Contents Safety Considerations ........................3 Regulatory Statement ........................6 Specifications ..........................7 Preface ............................9 Notices ............................9 Limited Warranty ........................... 10 Installation Requirements ......................11 Installation and Connecting Cables ....................12 Maintenance and Operation ......................13 Waste Treatment .......................... 13 1.

  • Page 3: Safety Considerations

    Safety Considerations To ensure operation safety, this instrument must be operated correctly and maintained according to a regular schedule. Carefully read to fully understand all safety precautions in this manual before operating the instrument. Please take a moment to understand what the signal words WARNING!, CAUTION and NOTE mean in this manual.
  • Page 4 WARNING! Do not operate the Monolith NT.115 with substances and under conditions which cause a risk of explosion, implosion or release of gases. Only operate the Monolith NT.115 with aqueous solutions. CAUTION Ensure that the power plug of the external power supply is easily accessible. The Monolith NT.115 instrument has to be installed in a way that it does not hinder the access to the external...
  • Page 5 CAUTION Only open the sample loading site when moving parts inside the instrument are at rest. Do not insert or remove a sample while linear actuator, laser or LED is at work. Moving parts in the instrument can be harmful. CAUTION Do not use the instrument in a cold room.

  • Page 6: Regulatory Statement

    Regulatory Statement The following safety and electromagnetic standards were considered: • IEC 61010-1:2001 Safety requirements for electrical equipment for measurement, control and laboratory use. Part 1 General Requirements • IEC 61010-2-010:2005 Safety requirements for electrical equipment for measurement, control and laboratory use. Part 2-010: Particular requirements for laboratory equipment for the heating of materials.

  • Page 7: Specifications

    Wavelength: 1475 nm +/- 15 nm Power: 120 mW max. Monolith NT.115 laser classification Device is LASER PRODUCT CLASS 1 Temperature control Range: 22°C – 45°C (at Room Temperature 25°C) Accuracy: +/- 0.3°C Noise level of Monolith NT.115 instrument: max. 64 dB(A)
  • Page 8 Connections All ingoing and outgoing connections can be found at the rear panel of the instrument. Name Function Ethernet Connector for connecting the Ethernet cable to the PC/notebook. Connector for connecting the USB cable to the PC/notebook. Only used for service by NanoTemper Technologies service. When the switch is turned on (position “I”), the instrument is Power Switch switched on.

  • Page 9: Preface

    Preface This manual is your guide for using the Monolith NT.115 and doing MicroScale Thermophoresis (MST) measurements. It instructs first-time users on how to use the instrument, and serves as a reference for experienced users. Before using the Monolith NT.115 instrument, please read this instruction manual carefully, and make sure that the contents are fully understood.

  • Page 10: Limited Warranty

    Limited Warranty Products sold by NanoTemper Technologies, unless otherwise specified, are warrantied for a period of one year from the date of shipment to be free of defects in materials and workmanship. If any defects in the product are found during this warranty period, NanoTemper Technologies will repair or replace the defective part(s) or product free of charge.

  • Page 11: Installation Requirements

    (RH), condensation may deteriorate optical components. Operate the instrument in an atmospheric pressure range of 800 – 1060 hPa. 10. Do not operate the Monolith NT.115 under conditions which pose a risk of explosion, implosion or the release of gases.

  • Page 12: Installation And Connecting Cables

    Connecting the power supply and the Monolith NT.115 Confirm that the power switch of the Monolith NT.115 instrument is off (power switch is at the back, left of the instrument).

  • Page 13: Maintenance And Operation

    Malfunction In case of a malfunction switch off the Monolith NT.115 instrument and wait for five minutes, then switch on the instrument again. If the malfunction persists switch off the device, unplug the power cable and contact the NanoTemper Technologies customer support.

  • Page 14: Introduction To Microscale Thermophoresis (Mst)

    1. Introduction to MicroScale Thermophoresis (MST) MicroScale Thermophoresis (MST) provides an easy, fast and precise approach to quantify biomolecular interactions. It allows for the detection of a wide range of interaction types, ranging from ion and fragment binding up to interactions of macromolecular complexes such as liposomes or ribosomes [1-3].
  • Page 15 In some cases, the fluorescence of the target molecule changes upon ligand binding even in the absence of an IR-induced temperature gradient. These cases can be analyzed using the Fluorescence Analysis Mode. The phenomenon is referred to as ligand-induced fluorescence quenching or enhancement and is typically caused by changes in target conformation or binding of the ligand in proximity to the target’s fluorophore(s).
  • Page 16 where [A] is the concentration of free fluorescent molecule, [L] the concentration of free ligand and [AL] is the concentration of the complex of A and L. The free concentrations of A and L are [A]=[A [AL] and [L]=[L ]-[AL], respectively. [A ] is the known concentration of the fluorescent molecule and ] is the known concentration of added ligand.

  • Page 17: Literature

    1.1.4. Software MST experiments are analyzed using the dedicated MO.Control and MO.Affinity Analysis Software, using the above equations. Prior to determining binding affinities, the MO.Control software performs several automated quality checks of the samples to detect protein adsorption, aggregation, or other irregularities. Based on the results, a guided assay development algorithm provides detailed instructions on how to efficiently improve assay quality (if necessary).

  • Page 18: Monolith ® Nt.115 Instrument

    = 1 nM - 500 mM). 2.3. Sensitivity The Monolith NT.115 can measure as little as the binding of single ions (40 Da) or small molecules (300 Da) to a target as well as the binding of large complexes such as ribosomes (2.5 MDa).

  • Page 19: Capillary Format

    2.5. Capillary Format The capillary format used for the Monolith NT.115 is inexpensive, easy to handle and offers maximum flexibility in the experiment scale. The standard sample tray format allows automatic processing of up to 16 capillaries (e.g. for a detailed K analysis).

  • Page 20: Preparation Of Mst Experiments

    3.1. Control Kits - Performing Standard Experiments Control Kits containing standardized sample material are recommended when using the Monolith NT.115 instrument for the first time for monitoring the correct performance of the Monolith NT.115 for training of new laboratory members Optimized Control Kits for use in MicroScale Thermophoresis are available either with red- or green- fluorescently labeled biomolecules.

  • Page 21: Fluorescence Labeling

    3.3. Fluorescence Labeling The following kits offer optimized fluorescence labeling and purification protocols for use in MicroScale Thermophoresis. The dyes have been widely tested with MST and the protocols are optimized to remove free, non-reacted dye: Cat # Protein Labeling Kits Reactions Monolith L001...
  • Page 22 Place the sample tray in the instrument by pushing it into the tray slot of the instrument as far as possible. The Monolith NT.115 instrument scans the tray and automatically determines the position of the capillaries on the tray.

  • Page 23: Using The Monolith Nt.115 Instrument

    4. Using the Monolith NT.115 Instrument Turn on the Monolith NT.115 instrument using the power toggle switch on the back of the instrument and start the control notebook. The display of the Monolith NT.115 instrument will show the following screen: The words “Ready”...
  • Page 24 Contact NanoTemper Technologies GmbH Floessergasse 4 81369 Munich Germany Phone: +49 (0)89 4522895 0 Fax: +49 (0)89 4522895 60 info@nanotempertech.com http://www.nanotempertech.com MicroScale Thermophoresis™ is a trademark. ® ® NanoTemper and Monolith are registered trademarks. ® ® NanoTemper and Monolith are registered in the U.S. Patent and Trademark Office.

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