Thermal Shift Analysis

Thermal shift analysis (TSA) is a thermal denaturation assay for elucidating optimal conditions for protein stabilization in solution. Protein destabilization and melting is measured as hydrophobic binding sites within the protein become exposed during unfolding and are able to bind a fluorescent dye. The melting temperature of a protein (Tm) is identified as the temperature at which 50% of the protein is unfolded.  The assay is performed in a 96 well plate format, allowing many conditions (buffers, ligands, etc.) to be screened simultaneously.

Available thermal shift assays:

• Tm determination for batch to batch validation
• Buffer screening for formulation optimization
• Ligand binding studies
• Protein:protein binding studies
• Construct stability comparisons

To order this service, click here or on the order tab at the top of the page.

Results of a thermal shift assay for a representative protein formulated in a series of buffers that range in pH from 5-9.5.  The left panel represents the raw fluorescence data and the right panel shows to the first derivative curves.  The peak minimum of the first derivative curves corresponds to the protein melting temperature.  For the example shown, the stability of the protein increases with decreasing pH.