Using High Content Imaging to Quantify Target Engagement in Adherent Cells
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Updated: Jan 31, 2026

Using High Content Imaging to Quantify Target Engagement in Adherent Cells
Published on: November 29, 2018
Hanna Axelsson1, Helena Almqvist1, Brinton Seashore-Ludlow2
1Chemical Biology Consortium Sweden, Science for Life Laboratory, Karolinska Institutet, Solna; Department of Medical Biochemistry and Biophysics, Karolinska Institutet, Solna.
Abstract:
Quantitating the interaction of small molecules with their intended protein target is critical for drug development, target validation and chemical probe validation. Methods that measure this phenomenon without modification of the protein target or small molecule are particularly valuable though technically challenging. The cellular thermal shift assay (CETSA) is one technique to monitor target engagement in living cells. Here, we describe an adaptation of the original CETSA protocol, which allows for high throughput measurements while retaining subcellular localization at the single cell level. We believe this protocol offers important advances to the application of CETSA for in-depth characterization of compound-target interaction, especially in heterogeneous populations of cells.
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