Using High Content Imaging to Quantify Target Engagement in Adherent Cells.

Axelsson H, Almqvist H, Seashore-Ludlow B

J Vis Exp - (141) - [2018-11-29; online 2018-11-29]

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.

Chemical Biology Consortium Sweden (CBCS) [Technology development]

PubMed 30582591

DOI 10.3791/58670

Crossref 10.3791/58670

Publications 9.5.0