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FRENCH INFRASTRUCTURE FOR INTEGRATED STRUCTURAL BIOLOGY

Thermal-Shift-Assay.jpg

INSTRUCT CENTER - FRANCE 1 - IGBMC

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)