Differential Scanning Fluorimetry / Thermal Shift Assays

Differential Scanning Fluorescence (DSF), utilises a fluorescent dye, e.g. SYPRO Orange, and a qPCR machine.  The dye binds non-specifically to hydrophobic surfaces on the protein. The dye is quenched by water. The sample mix is heated, and the protein unfolds, exposing the hydrophobic core, which leads to an increase in fluorescent signal. The fluorescent signal is plotted against temperature, and the melting temperature, Tm, of the protein can be calculated. By comparing the Tm in a range of different buffer conditions, we can find the conditions that make the protein the most stable. The method also allow high-throughput screening of ligands and small molecules by keeping the conditions identical, and look for increased stability upon addition of ligand.

We have an initial in-house screen that will screen a wide range of pH and salt concentrations, but we are also able to run bespoke screens.

Please contact June Southall.