Bruker AVANCE NEO - 500 MHz
Applications
Analysis of small molecules, peptides and biomolecules of small molecular weight. Biomolecule (protein, RNA, DNA) – small molecule titrations. Rapid assessment of the quality of a biomolecular sample in order to initiate biophysical studies.
NMR tubes : 3 or 5 mm in diameter (standard or Shigemi)
Temperature : 0 – 80 ˚C
Magnet and console
Shielded, superconducting Oxford magnet of 500 MHz (11.7 Tesla) coupled to a Bruker AVANCE NEO console.
Probes available
Normally equipped with a 5 mm TXI probe (inverse triple resonance) with automatic tuning and matching and Z-gradient capability. This probe is designed for 1H observation with decoupling capacity of 13C, 15N and 2D.
A second probe is available if needed: a 5 mm QXI probe (inverse quadruple resonance) with automatic tuning and matching and Z-gradient capability. This probe is designed for 1H observation with decoupling capacity of 13C, 15N and 31P in the same experiment. Can also be used for 13C, 15N and 31P observed experiments, albeit not optimized for that purpose.
Software
- TopSpin 4.1.3 (Linux CentOS)
- Complete Molecular Confidence suite (CMC-assist and CMC-se)
- Non-Uniform Sampling (NUS)
- Protein Dynamics Center
Bruker AVANCE NEO - 600 MHz with cryo-probe
Applications
Improved sensitivity with cryo-probe usage. Ideal for multi-dimensional, triple resonance experiments typically used in structural and dynamics studies of proteins and nucleic acids.
NMR tubes : 3 or 5 mm in diameter (standard or Shigemi) and Bruker shaped tubes (with SPU)0
Temperature : 0 – 80 ˚C
Magnet and console
Superconducting Oxford magnet of 600 MHz (14.1 Tesla) coupled to a Bruker AVANCE NEO console.
Probes available
Normally equipped with a TCI 5 mm cryo-probe (inverse triple resonance) with automatic tuning and matching, Z-gradient capability and sample positioning unit (SPU). This probe is designed for high sensitivity 1H observation (with 13C, 15N and 2D decoupling) as well as 13C observation (with 1H, 15N and 2D decoupling). The pre-amplifier for the lock signal (2D) is also cryo-cooled thus enabling the usage of lower percentages of deuterated solvent.
A second probe is also available on demand: a 5 mm TXI probe (inverse triple resonance) with automatic tuning and matching and Z-gradient capability. This probe is designed for 1H observation with decoupling capacity of 13C, 15N and 2D.
Software
- TopSpin 4.1.3 (Linux CentOS)
- Complete Molecular Confidence suite (CMC-assist and CMC-se)
- Non-Uniform Sampling (NUS)
- Protein Dynamics Center
Bruker AVANCE NEO - 700 MHz
Applications
Great resolution and sensitivity. Ideal for multi-dimensional, triple resonance experiments typically used in structural and dynamics studies of proteins and nucleic acids.
NMR tubes : 3 or 5 mm in diameter (standard or Shigemi)
Temperature : 0 – 80 ˚C
Magnet and console
Shielded, superconducting Bruker Ascend magnet of 700 MHz (16.4 Tesla) coupled to a Bruker AVANCE NEO console.
Probes
Normally equipped with a 5 mm TXI probe (inverse triple resonance) with automatic tuning and matching and Z-gradient capability. This probe is designed for 1H observation with decoupling capacity 13C, 15N and 2D.
A second probe is also available on demand: a BBI 5 mm probe (inverse broadband) with automatic tuning and matching and Z-gradient capability. This probe enables the observation of 1H with decoupling capacity for a single nucleus between 31P and 109Ag depending on the tuning. Useful for HSQC, HMQC and HMBC experiments. Can be used for the observation of 13C, 15N or 31P observed experiments, albeit not optimized for that purpose.
Software
- TopSpin 4.1.3 (Linux CentOS)
- Complete Molecular Confidence suite (CMC-assist & CMC-se)
- Non-Uniform Sampling (NUS)
- Protein Dynamics Center
Small Angle X-rays Scattering (SAXS)
Small angle X-rays scattering is a biophysical method for the analysis of biomolecules (proteins, RNA, DNA) in solution and provides information regarding to variations in the electronic density of molecular structures in solution. SAXS yields information on both size, shape, density and molecular weight of particles.
Description of the equipment
The SAXSLAB Bio-Nordic is equipped with a vacuum GANESHA platform which enables the measurement of X-rays scattering at small angles for vectors of momentum transfer (q-values) from 0.0006 to 3.4 Å-1, for both SAXS (small angle) and WAXS (wide angle) applications, for particles up to 150 nm in diameter.
X-rays source
The X-rays are generated by a Excillum MetalJet D2+ 70 kV source using a 95 % gallium alloy (liquid jet anode – 1.34 Å). This powerful X-rays source offer superior brightness when compared to a conventional rotating anode and thus enables fast analysis of samples.
Size-exclusion chromatography system
The optional addition of an ÄKTAmicro chromatography system (GE Healthcare) enables size-exclusion type separation of biomolecules prior to SAXS analysis. The inline attachment lets the research to simultaneously separate by chromatography and analyze by SAXS each detected peak. A UV light/detector (Biocube, SAXSLAB) is also present on the SAXS system for direct absorbance measurement.
Detector
The Bio-Nordic is equipped with a Dectris hybrid photon counting detector (Dectris EIGER2 R 1M) offering micrometric spacial resolution (75 µm x 75 µm) and a wide dynamic range and a large pixel array (1M pixels). The detector is water-cooled and installed in a vacuum chamber therefore eliminating air scattering.
High-throughput screening
A high-throughput robot installed on the Bio-Nordic enables screening refrigerated 96-well plates by automating liquid handling. Less than 20 µl is required per sample.
Software
- Linux Ubuntu 16.04
- RAW 1.4.2 (BioXolver version)
- ATSAS 2.8.4
- DENSS
Size Exclusion Chromatography with Multi-Angle Light Scattering analysis (SEC-MALS)
Size Exclusion Chromatography with Multi-Angle Light Scattering analysis (SEC-MALS) is an absolute technique for the determination of the molar mass and the average radius of gyration (Rg) of biomolecules in solution following a separation by liquid chromatography. The technique uses static diffusion of a high intensity light source through a solution containing macromolecules. The intensity of this diffusion is then detected under several angles, hence being able to calculate precisely the molar mass and the average radius of gyration (Rg).
Description of the equipment
Analytical chromatography (SEC)
An ÄKTAmicro (GE Healthcare) liquid chromatography system is used to perform a high resolution separation of macromolecules (proteins, RNA, DNA) at analytical levels. The precision of low flow rate pumps enables the use of analytical-grade chromatography columns and injection of small quantities of material to be analyzed.
Multi-Angle Light Scattering (MALS) Detector
A Dawn HELEOS II (Wyatt Technology) multi-angle light scattering detector is connected directly at the outlet of the chromatography column. The presence of multiple light scattering detectors enables the measurement of the light intensity at 18 different angles, offering great sensitivity (as low as 200 ng of BSA in solution) and both a wide range of molecular weights (200 Da to 1000 MDa) and of size (Rg 10 to 500 nm).
Refractive Index Detector (RI)
Finally, a OptiLab T-rEX (Wyatt Technology) refractive index detector is positionned downstream of the MALS detector in order to determine the concentration of biomolecules being separated.
Available chromatography columns
Several analytical liquid chromatography columns are available for SEC-MALS analysis.
Size-exclusion columns (SEC)
- Superdex 200 Increase, 10×300
- Superose 12, 3.2×300
- Superdex 200 Increase, 3.2×300
- Superdex 75, 3.2×300
- Superose 6 Increase, 3.2×300
Ion-exchange columns (IEX)
- Mono S, 1.6×5
- Mono Q, 1.6×5
Software
- ASTRA 6.1.6.5 (Wyatt Technology)
Isothermal titration calorimetry
Isothermal titration calorimetry (ITC) is a biophysical technique used in the quantitative study of biomolecular interactions. The heat released or absorbed at the time of binding is measured directly, and allows for the simultaneous determination of all the thermodynamic parameters of the binding, as well as the affinity and the stoichiometry of the interaction. This measurement is carried out in solution, without prior modification of the partners.
Description of the equipment
The MicroCal iTC200 provides a detailed overview of the thermodynamic parameters of the binding reaction. The system has a 200 μl sample cell, and a 50 µl titration syringe, and allows direct measurement of the heat absorbed or released by mixing precise amounts of reagents during the experiment.
Data analysis is performed using Origin™ software, in which the user obtains the stoichiometry (n), dissociation constant (KD) and enthalpy (ΔH) of the interaction. The Origin software can also be used to fit more complex binding patterns (two sets of sites, competition binding etc.).
It is also possible to use the ITC control software to simulate an experiment and determine the optimal receptor and ligand concentrations to use.
Software
- MicroCal iTC200 Control Software
- OriginLab Origin 7.0
Varian Cary 300 – UV and visible
The UV Cary 300 is a spectrophotometer used for UV and visible light spectroscopy. The instrument can be used for absorbance measurement at fixed or variable wavelength, for enzyme kinetics studies or for heat denaturation curves.
Varian Cary Eclipse – Fluorescence
The Cary Eclipse is a spectrofluorometer used for fluorescence spectroscopy studies of biomolecules.