Thermal Shift Assay (TSA)

Thermal Shift Assay (TSA)


Thermal shift assay (TSA) is a technique to quantify the change of thermal denaturation temperature of proteins in different environments, which is widely used in the characterization of protein-ligand interactions, screening of mutants, buffers, detergents, etc. TSA not only detects whether a small molecule binds to the target protein or not, but also can be used in the identification of the target of the drug candidate and target binding. In addition, TSA can be used to screen out small molecule fragments that have weak interactions with target proteins, and then optimize the active fragments based on their structural information to obtain more active lead compounds for new drug development.

thermal shift assay (TSA) technologyFigure 1. Principle of TSA. (Petrauskas V, et al., 2023)

Working Principle

TSA detects the thermal stability of proteins using the fluorescent dye SYPRO Orange, an environmentally sensitive hydrophobic dye, as an indicator. When the temperature rises, the protein unfolds and the hydrophobic part is exposed. The dye then binds specifically to the hydrophobic portion of the protein and fluorescence is enhanced. When a specific compound binds to the target protein, it stabilizes the protein, thereby increasing the melting temperature.


  • Low protein sample loss.
  • High throughput and accurate data.
  • Wide range of temperature variation.

CD BioSciences has a comprehensive technology platform dedicated to providing thermal shift assay (TSA) services for drug screening, protein stability, and biomolecular interaction studies.

Main Applications

  • Drug Screening

TSA can be used to study drug interactions and conformational changes with target proteins during drug screening. We can provide TSA technology for the rapid determination of binding sites and thermal stabilization effects of drugs to assess potential efficacy and mechanism of action.

  • Protein Stability Analysis

Most proteins have stringent temperature requirements, and the assessment of protein thermal stability contributes to the normal expression of protein functions. We use TSA not only to determine the thermal stability parameters of proteins in high throughput but also to evaluate the effects of mutation, pH, ionic strength, and other factors on the thermal stability of proteins.

  • Biomolecular Interaction Analysis

TSA can be used to study the interactions and binding sites between biomolecules, such as DNA-proteins, proteins-ligands, and enzymes-substrates. We provide TSA technology to quickly analyze the binding constants and thermodynamic parameters between biomolecules, which helps us to deeply understand the interaction mechanism of biomolecules.

What Can We Offer?

CD BioSciences possesses a wealth of resources devoted to providing expert thermal shift assay (TSA) technical support to our customers. If you are interested in our services, please feel free to contact us.


  1. Petrauskas V, Kazlauskas E, Gedgaudas M, et al. (2023). Thermal shift assay for protein–ligand dissociation constant determination[J]. TrAC Trends in Analytical Chemistry. 117417.
For research use only, not intended for any clinical use.
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