Kit for fluorescence-based measurement of protein unfolding or thermal stability in thermal shift assay, also called Protein Thermal Shift™, differential scanning fluorimetry, or Thermofluor assay.
Product Description
The GloMelt™ Thermal Shift Protein Stability Kit is used to detect protein unfolding or measure thermal stability by performing a thermal shift assay, also called Protein Thermal Shift™, differential scanning fluorimetry, or Thermofluor assay.
Features
Green fluorescence optimal for qPCR instruments and ROX normalization
Compatible with high detergent concentrations
Tolerates wide pH range, reducing agents, and common buffers/excipients
Great for high-throughput assays, low reaction volumes, and low protein concentrations
Highly soluble and stable in aqueous buffers
Kit Components
GloMelt™ Dye, 200X
Goat IgG Control, 10 mg/mL
ROX Reference Dye, 40 uM (33022-T/33022-1 only)
Spectral Properties
GloMelt™ Dye Ex/Em 468/507 nm
ROX Reference Dye Ex/Em 575/600 nm
About GloMelt™ Thermal Shift Protein Stability Kit
Environmentally sensitive fluorescent dyes can be used to monitor the temperature dependent unfolding of a protein. The protein’s melting temperature (Tm) is a reporter of the protein’s thermal stability.
GloMelt™ dye undergoes fluorescence enhancement upon binding to hydrophobic regions of denatured proteins, and therefore can be used to detect protein unfolding or measure thermal stability by performing a thermal shift assay, also called Protein Thermal Shift™, differential scanning fluorimetry, or Thermofluor assay.
The thermal shift assay is a rapid and inexpensive technique that quantifies change in protein denaturation temperature, and thus can be used to screen conditions that affect protein thermal stability, such as protein mutations, ligand binding, and buffer formulations (like pH, salts, detergents, and other additives). These assays are rapid (typically about 30 minutes) and are performed on a quantitative PCR system. The thermal shift method is compatible with high throughput screening and requires much less protein than methods such as circular dichroism and differential scanning calorimetry.
To learn more about GoMelt™ and see how it compares to competitor products, visit our GoMelt™ Technology Page.
Possible Uses for Thermal Shift Assays
Optimize buffer formulation for protein stability and storage
Determine how mutations affect your protein’s stability
Rapidly screen small molecule drug candidates and other ligands for protein binding
Small molecule inhibitors of bromodomain proteins have shown therapeutic effects in cancer models. Here, a GloMelt™ thermal shift assay was performed on 10 ug bromodomain BRD2 in the presence of bromodomain inhibitors JQ1 or RVX-208. Inhibitor binding stabilized the protein as indicated by the shift of the melting curves.
GloMelt™ Dye vs. Competitors
GloMelt™ dye has significant advantages over other environmentally sensitive dyes, such as SYPRO® Orange and PROTEOSTAT® TS dye. GloMelt™ dye generates a strong signal because it is optimized for detection in the SYBR® Green channel of qPCR instruments, and therefore low reaction volumes and low protein concentrations can be used. GloMelt™ dye is compatible with high concentrations of protein stabilizers (such as glycerol and sorbitol), and also protein destabilizers (such as DTT and imidazole). GloMelt™ dye performs very well in high detergent concentrations, unlike SYPRO® Orange. Another advantage is that ROX dye can be included with GloMelt™ dye during thermal shift assays, which improves results by increasing replicate consistency in PCR instruments that require ROX passive reference dye.
SYBR is a registered trademark of Thermo Fisher Scientific. Proteostat is a registered trademark of Enzo Life Sciences.
This website uses cookies so that we can provide you with the best user experience possible. Cookie information is stored in your browser and performs functions such as recognising you when you return to our website and helping our team to understand which sections of the website you find most interesting and useful.
Strictly Necessary Cookies
Strictly Necessary Cookie should be enabled at all times so that we can save your preferences for cookie settings.
If you disable this cookie, we will not be able to save your preferences. This means that every time you visit this website you will need to enable or disable cookies again.