Discovery Biology, Discovery Sciences, IMED Biotech Unit, AstraZeneca, Cambridge, UK.
Methods Mol Biol. 2021;2263:201-215. doi: 10.1007/978-1-0716-1197-5_8.
A thermal shift assay (TSA) involves measuring the effect of a compound on the thermal stability of a protein as an indirect measure of ligand binding. In this chapter, we provide a protocol for a conventional TSA with recombinant/purified proteins using differential scanning fluorimetry (DSF), followed by a protocol for a Cellular Thermal Shift Assay (CETSA), which measures the soluble cellular protein remaining after a transient heat shock of live cells to detect intracellular ligand binding.
热迁移分析（TSA）涉及测量化合物对蛋白质热稳定性的影响，作为配体结合的间接测量。在本章中，我们提供了使用差示扫描荧光法（DSF）对重组/纯化蛋白进行常规 TSA 的方案，随后提供了细胞热迁移分析（CETSA）的方案，该方案测量活细胞短暂热休克后可溶性细胞蛋白的剩余量，以检测细胞内配体结合。
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