Main Content
Core Facility Equipment
- ÄKTA (Protein purification)
- Prometheus NT.48 (Differential Scanning Fluorimetry - DSF)
- Bruker EMX-6/1 (EPR spectroscopy)
- Jasco J-815 (CD spectroscopy)
- Biologic SFM-20 (Stopped flow kinetics)
- Monolith NT.115 (Microscale Thermophoresis - MST)
- Octet K2 (Bio-Layer Interferometry - BLI)
- Jasco FP-6300 (Fluorescence spectroscopy)
- Complementing equipment
ÄKTA
Protein purification
ÄKTA purifier systems (GE Healthcare), and a variety of chromatography columns are available. The systems are equipped with flexible fraction collectors and multiple-wavelength detectors, all contained in refrigerated or anaerobic cabinets.
Prometheus NT.48
Differential Scanning Fluorimetry
A Nanotemper Prometheus NT.48 (nanoDSF) system allows label-free analysis of thermal and chemical protein stability in small-sized samples by measuring internal tryptophane fluorescence.
Bruker EMX-6/1
EPR spectroscopy
X-Band-EPR spectrometer (Bruker EMX-6/1) composed of EMX113 console, ER-041 XG X-band microwave bridge with ER-041-1161 microwave frequency counter and EMX-1101 magnet (11-640 mT). A standard-Universal TE102 rectangular (ER-4102) and a dual-mode cavity are available. Measurements are usually performed at liquid nitrogen temperature. The use of an Oxford instruments variable temperature Helium flow cryostat (4-200 K) is possible. For expert users only; available on request.
Jasco J-815 CD
CD spectroscopy
A Jasco J-815 CD spectrometer purged with argon allows to determine the content of secondary structural elements in the UV range. With the programmable PTC-423S Peltier-element thermostat melting curves can be recorded. The superb sensitivity of the Jasco machine makes time-dependent CD spectroscopyin the visible range for heme or [2Fe-2S] cluster-containing proteins possible.
Biologic SFM-20
Stopped flow kinetics
The SFM-20 stopped-flow system is routinely set up for spectroscopic detection of rapid kinetic processes. The system can be swapped to UV-Vis, fluorimetric and, if sensitivity allows, CD polarimetric detection. Since the removable syringes are air-tight and anaerobic tents are in the same building, we have the possibility to anaerobically study kinetic processes.
Monolith NT.115
Microscale Thermophoresis (MST)
U sing a Nanotemper Technologies Monolith NT.115 microscale thermophoresis instrument, complex formation between a fluorescently labelled protein and another protein or ligand can be detected with minimal effort requiring limited quantities of protein and/or ligand.
Octet K2
Bio-Layer Interferometry (BLI)
The Pall ForteBio Octet K2 system allows the label-free analysis of biomolecular interactions between an immobilized compound at the tip of a biosensor and an analyte in solution. Binding assays enable the determination of complex formation, its kinetics, and the underlying affinities.
Jasco FP-6300
Fluorescence spectroscopy
A Jasco FP-6300 fluorescence spectrometer is avalaible with computer interface for kinetics and scanning. For large numbers of samples the Tecan Infinite M200 plate reader, albeit of lower resolution, can be used.
Complementing equipment
Complementary to the core facility equipment the Institute of Cytobiology maintains infrastructure which is available on request:
Sample preparation
An EmulsiFlex-C3 high-pressure homogenizer allows the disruption of pro- and eukaroytic cells at large-scale and facilitates the preparation of cell extracts.
Anaerobic facilities
Two Coy anaerobic tents (2-5 % hydrogen in nitrogen) can be used for aqueous biochemistry particularly of air-sensitive (metallo-) proteins. Substantial expertise is available to support respective challenging experiments, and spectrometers on site allow immediate analyses.
UV-Vis spectroscopy
In addition to the equipment of the core facility several UV-Vis spectrometers are hosted by our institute: a Jasco V-550 interfaced with external computer, two Kontron Uvikonspectrometers (922 and 933, internal computer), and a Nanodrop spectrometer.
HPLC
Analytical HPLC (Waters/Raytest) may be used to trace molecules of interest by absorption or radioactivity.
Luminescence
A Berthold Sirius L – tube luminometer is the tool of choice for high-sensitivity luminescence measurements.
Multiwell Reader
Tecan Infinite plate readers are suitable for absorption, fluorescence, luminescence, and fluorescence polarisation.
Flow cytometry
A BD FACSCanto II flow cytometer enables the realisation of cell-based assays.