= Differential scanning fluorimetry
<Differential scanning fluorimetry> follows <fluorescence> while a <protein> sample undergoes controlled heating. Changes associated with <protein> unfolding provide a transition curve and an <apparent protein unfolding midpoint>. Comparing matched conditions turns the method into a <thermal shift assay>.<Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery by Mei-Chu Lo and others>[Abstract; temperature-controlled fluorescence measurement]{ref}
In a common implementation, an environment-sensitive fluorescent dye reports changes as normally buried regions become exposed. Compare otherwise matched <protein>-plus-dye samples with and without a candidate <ligand>, rather than comparing measurements made with different buffers or heating programs. Fluorescent compound-only and dye-only controls can help diagnose reporter interference; these are proposed controls, not specific settings claimed for every published assay.
For an illustrative result, midpoints of $320\,\mathrm{K}$ and $324\,\mathrm{K}$ give a thermal shift of $+4\,\mathrm{K}$. The measured quantity is a change in a heating-dependent transition. An independent <ligand binding> or activity measurement is needed to assess the proposed molecular mechanism, as illustrated by the comparisons with <isothermal titration calorimetry> in the 2004 evaluation.<Evaluation of fluorescence-based thermal shift assays for hit identification in drug discovery by Mei-Chu Lo and others>[Abstract; orthogonal comparisons and analysis pitfalls]{ref}
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