Differential scanning fluorimetry
Description
DSF is a spectrophotometric technique that monitors thermally induced protein unfolding. This method utilizes dyes that fluoresce under non-polar environments, including exposed hydrophobic regions of the protein structure. As the protein is heated and denatures, hydrophobic regions will be exposed increasing the fluorescence of the sample which can be monitored using real-time PCR machines. The temperature at half of the maximum fluorescence is defined as the protein melting temperature.
Acknowledgements
References
Niesen, F., Berglund, H. & Vedadi, M. The use of differential scanning fluorimetry to detect ligand interactions that promote protein stability. Nat Protoc 2, 2212–2221 (2007). https://doi.org/10.1038/nprot.2007.321
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