Thermal Shift Assay (Differential Scanning Fluorimetry) Service
Thermal shift assays utilize differential scanning fluorimetry (DSF) to measure the thermal stability of a target protein and the increase in protein melting temperature upon the binding of a stabilizing agent. It is useful for identifying ligands, buffer conditions, and cofactors affecting protein stability.
Thermal shift assays are performed via qPCR for high-throughput data acquisition in the 384 well format. We offer thermal shift assays for reader domain proteins, including bromodomains, chromodomains, and other methyl-readers. In addition, we offer custom-tailored thermal stability assay development for many other target categories.
Upon binding compounds cause a considerable change in the melting temperature of the target protein. These assays are carried out by our well-versed scientists with completion of standard studies within two weeks.
- The protein thermal shift assay is used to identify ligands that bind to the target, buffer conditions, and cofactors
- The differential scanning fluorimetry technique requires minimal assay establishment and is very cost-effective
- Deliverable: melting temperature shift (∆Tm) as a readout for compound binding affinity
Thermal shift assay uses the phenomenon in which ligand binding increases the target protein's heat stability; this phenomenon may be utilized to gain insight in the compound-target interaction. Reach out to our business development team today to inquire about our differential scanning fluorimetry services.
Thermal Shift Assay Principle
The thermal shift assay measures the melting temperature of a protein (Tm), which is the temperature at which there is 50% denaturation. Protein denaturation is monitored via an increase in fluorescence of SYPRO Orange dye which binds to hydrophobic residues that get exposed as the target protein unfolds. Compound binding to the target protein leads to increased stability and an increase in melting temperature.
Thermal Shift Assay Details
Example of Differential Scanning Fluorimetry of cGAS in the presence or absence of 10 µM PF-06928215.
The thermal denaturation of cGAS is detected via SYPRO Orange dye. The addition of the inhibitor PF-06928215 stabilizes the protein and increases the melting temperature from 45 to 53 degrees Celsius.
Differential Scanning Fluorimetry of BRD2-Tndm in the presence or absence of (+)- JQ1. The thermal denaturation of BRDT-Tndm is detected via SYPRO Orange dye. Addition of the BET Bromodomain inhibitor/ligand (+)- JQ1 stabilizes the protein folding and shifts the Tm from 41 to 44 degrees Celsius.
Instruments: BCFX384 (BioRad) real-time PCR detection system
Turnaround time: The study is typically completed within 2 weeks after receipt of materials.
Projects are performed on a first-come-first-serve basis.
Sample requirements: Protein purity of 85% or more is required. The protein amount is project dependent. Typical usage is between 1-3 ug per reaction.
Please see our FAQ for shipping instructions.
Screening location: The assay is performed in Malvern, PA, USA.
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