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![[Figure 4]](qt5002fig4mag.jpg)
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Figure 4
The ThermoAffinity pipeline consists of three steps. In the first step, the data are loaded and preprocessed. The preprocessing includes smoothening or normalizing the data, adding information about the ligand concentration and selecting the `hot' and `cold' regions to obtain the normalized fluorescence Fnorm = Fhot/Fcold. The fluorescence signal versus time plot will be colour-coded using a base-10 log scale and the viridis palette. Secondly, the thermophoresis signal versus binding partner concentration curve, together with the protein concentration, is used to obtain the binding affinity (Kd) and the numeric values RF1 and RF2 (the fluorescence signal of 1 µM unbound protein and 1 µM complexed protein, respectively). All of the estimated parameters are presented with their respective errors and associated confidence intervals. Finally, individual CSV files with the model-derived information can be exported. |
![[Figure 4]](qt5002fig4.jpg)
 | STRUCTURAL BIOLOGY |
ISSN: 2059-7983
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