This technology measures the thermal stability of proteins by following the fluorescence of a fluorescent dye, the orange SYPRO. A thermal ramp (20 to 95°C) is applied and during protein denaturation hydrophobic areas will appear, the orange SYPRO will attach itself to it and its fluorescence will increase.
To perform these measurements we use the StepOne plus (Applied Biosystem) device.
These rapid measurements (about 1h), carried out in a plate of 96 wells, make it possible to carry out a stability screening of the protein in different buffers or in the presence of different ligands. This technique uses very little material: about 20 µl/ protein well at 0.1 -0.5mg/ml.