Related Experiment Video
Updated: Aug 26, 2025

Assessing Cellular Target Engagement by SHP2 PTPN11 Phosphatase Inhibitors
Published on: July 17, 2020
Two-dimensional electrophoresis-cellular thermal shift assay (2DE-CETSA) for target identification of bioactive
Makoto Muroi1, Hiroyuki Osada2
1Biomolecular Characterization Unit., Technology Platform Division, RIKEN Center for Sustainable Resource Science, Saitama, Japan; Chemical Resource Development Unit, Technology Platform Division, RIKEN Center for Sustainable Resource Science, Saitama, Japan.
Abstract:
Identification of target molecules of new bioactive compounds is still a challenge in drug development. Various proteomics-based methods have been developed to analyze the interaction between compounds and target proteins. Among these methods, cellular thermal shift assay (CETSA) has been frequently applied in recent years for validation studies of compound-protein interactions using antibodies. Combining CETSA with comprehensive proteomic analysis has been successful in narrowing down the target(s) of a new compound from the enormous number of proteins in cell. In this chapter, we introduce 2DE-CETSA, which combines CETSA with proteome analysis using two-dimensional electrophoresis as a method for identification of target proteins.
Related Concept Videos
Two-dimensional Gel Electrophoresis
The first dimension separation uses the isoelectric focusing or IEF technique performed on immobilized pH gradient (IPG) strips that separate proteins according to their isoelectric points.
Biological samples, such...
Electrophoresis: Overview
There...
Capillary Electrophoresis: Applications
Capillary zone electrophoresis (CZE) separates ionic components based on their electrophoretic mobility. It has been used to separate proteins, amino acids,...
Western Blotting
The technique begins with separating proteins from the sample using sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), followed by protein transfer, immunoblotting, and finally, protein detection.

