Monitoring changes in the melting temperature of a target protein (i.e., thermal shift assay, TSA) is an efficient method for screening fragment libraries of a few hundred compounds. We present a TSA protocol implementing robotics-assisted nano-Differential Scanning Fluorimetry (nano-DSF) for monitoring intrinsic tryptophan fluorescence and light back-scattering for fragment screening.
Ahmad, M. U. D., Fish, A., Molenaar, J., Sreeramulu, S., Richter, C., Altincekic, N., Schwalbe, H., Wienk, H., Perrakis, A. Nano-Differential Scanning Fluorimetry for Screening in Fragment-based Lead Discovery. J. Vis. Exp. (171), e62469, doi:10.3791/62469 (2021).