Instruct-centre France-1 offers state-of-the-art equipment and expertise to perform quality control on protein samples.
System components: Multi-angle light scattering detector: miniDAWN TREOS from Wyatt Technology. Equipped with the QELS module which enables measurement of the hydrodynamic radius. Differential Refractive Index (dRI) detector: Optilab T-rEX from Wyatt Technology. Chromatography system: Ettan MicroLC from GE Healthcare.
Device: Thermofluor (use of extrinsic fluorescent dye to label the proteins) and Prometheus (no dye, native intrinsic fluorescence from the proteins).
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. Indeed, 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 optimization of proteins solubility and stability properties improvesthe 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 characterization and to assign functions.
Device: Thermofluor (use of extrinsic fluorescent dye to label the proteins) and Prometheus (no dye, native intrinsic fluorescence from the proteins)