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User Guide

Analytical ultracentrifugation (AUC)

AUC allows the characterisation of macromolecules and macromolecular self- and hetero-association processes in solution. Two types of complementary experiments, sedimentation velocity and sedimentation equilibrium, can be performed in an analytical ultracentrifuge which is a high-speed centrifuge equipped with an optical detection system. The observation of macromolecules or macromolecular complexes sedimentation gives access to their hydrodynamic and thermodynamic properties, including their size, shape, molar mass, degree of heterogeneity, oligomeric state, stoichiometry, and binding constants.

Isothermal titration calorimetry (ITC) 

ITC is used to investigate all types of protein interactions, including protein-protein interactions, protein-DNA/RNA interactions and protein-small molecule interactions.

Microscale Thermophoresis (MST)

MST is a powerful new technology to quantify biomolecular interactions in a few microliter solution. MST measures changes of mobility of the molecules in microscopic temperature gradients to determine binding affinities. 

This technology has several advantages over other standard techniques to analyze interactions, such as surface plasmon resonance (SPR) and isothermal microcalorimetry (ITC). It can measure affinities in free solution without surface immobilisation with low sample consumption and within sub-nM to mM range.  Experiments can be carried out with a broad range of solution conditions, including detergent mixtures and complex bioliquids. MST can monitor the binding of single ions (40Da) or small molecules (300Da) to a target as well as the binding of ribosomes (2.5MDa).

MALS

Size exclusion chromatography (SEC) coupled with multiangle light scattering (MALS) is a straightforward technique to determine the accurate molar mass and the average size of proteins and macromolecular complexes in solution. MALS can measure the absolute molar mass and size of molecules without the use of reference standards.

One of the major application is the determination of the size and stoichiometry of tightly bound heterocomplexes, such as protein/protein, protein/DNA, protein/RNA and protein/detergent interactions.

Thermal Shift Assay

Thermal shift assay is a thermodenaturation assay to monitor the thermal stability of proteins and investigate factors affecting this stability. This rapid and simple technique is used in high-throughputmode to screen optimalbuffer conditions, ligands, cofactors and drugs for purified proteins. Two methods to monitor protein denaturation are available: a differential scanning fluorimetry (DSF) method and a differential static light scattering method (DSLS).

The optimisation of proteins solubility and stability properties improves the success rate of their structural studies. Changes in the thermal stability of the protein–ligand or protein-peptide complexes relative to the stability of the protein alone allow to rapidly identify promising complexes for further structural characterisation and to assign functions.

Flow-Induced Dispersion Analysis (FIDA)

FIDA combines Taylor dispersion analysis (TDA) and fluorescence detection in straight fused silica capillaries to measure biomolecular size, enabling accurate determination of hydrodynamic radius (Rh) and size-based characterization of biomolecular interactions.

BLI Bio Layer Interferometry

Bio-Layer Interferometry (BLI) is a label-free technology for measuring biomolecular interactions. It is a fast and high throughput technique that enables real-time analysis for determination of affinity, kinetics and concentration.