In this context, we herein report new STAT3 binding assays based on differential scanning fluorimetry (DSF) and differential scanning light scattering (DSLS) to characterize interactions between STAT3 protein and inhibitors. Several peptide and small molecule STAT3 inhibitors have been evaluated, and new...
Here we present a set of measurements using Differential Scanning Fluorimetry (DSF) as an inexpensive, high throughput screening method to investigate the folding of silk protein molecules as they abandon their first native melt conformation, dehydrate a
References: Pooria Gill, et al. Differential scanning calorimetry techniques: applications in biology and nanoscience. Journal of Biomolecular Techniques. 2010 Dec; 21 (4): 167-93. Related Reading Nano Differential Scanning Fluorimetry (NanoDSF) Protein Thermal Shift Assay (PTSA) Thermal Gravimetric...
Here, we used differential scanning fluorimetry (DSF) to characterize the stability and ligand binding properties of this antibody and its cocaine-binding Fab fragment. The Sypro orange dye commonly used for DSF revealed multiple overlapping thermal protein denaturation transitions for both the mAb and...
A differential scanning fluorimetry (DSF) based high-throughput screening assay with the fluorescent molecular rotor CCVJ (9-(2-carboxy-2-cyanovinyl)julolidine) was developed. CCVJ is mainly sensitive to viscosity and less to polarity in comparison to polarity-sensitive dyes like SYPRO Orange, ...
1 .thermofluor Thermofluor (aka Differential Scanning Fluorimetry) 2 .thermofluor Thermofluor • Thermofluor gives an optical readout of protein melting as e.g. observed with DSC. • Optical readout is much more sensitive than direct thermal measurement. • TF consequently allows parallel measurem...
Differential scanning fluorimetry (DSF) is a widely used biophysical technique with applications to drug discovery and protein biochemistry. DSF experiments are commonly performed in commercial real-time polymerase chain reaction (qPCR) thermal cyclers or nanoDSF instruments. Here, we report the construct...
Differential scanning fluorimetry (DSF) is a widely used thermal shift assay for measuring protein stability and protein-ligand interactions that are simple, cheap, and amenable to high throughput. However, data analysis remains a challenge, requiring improved methods. Here, the program SimpleDSFviewer...
Theory and applications of differential scanning fluorimetry in early-stage drug discoveryThermal stabilityFoldingUnfoldingRefoldingFluorimetryLigands screeningCrystallizationBuffer optimizationTypical thermal denaturation profile of a protein sample. Fluorescence emission changes with the temperature. The sigmoidal curve...
Differential scanning fluorimetry is useful for a wide variety of applications including characterization of protein function, structure鈥揳ctivity relationships, drug screening, and optimization of buffer conditions for protein purification, enzyme activity, and crystallization. A limitation of classic ...