Principles
Differential Scanning Fluorimetry (DSF) evaluates protein stability by monitoring fluorescence signal changes during protein thermal denaturation. Common dyes like SYPRO Orange fluoresce upon binding exposed hydrophobic regions during denaturation, recording melting curves.
Core Applications
- Drug screening: Compare Tm shifts (ΔTm) with/without ligand for rapid binding assessment
- Condition optimization: Screen optimal buffer pH, salt concentration, additives
- Stability comparison: Compare stability of different mutants or modified forms
- Formulation development: Evaluate long-term stability under different formulation conditions
Advantages
- Extremely low sample requirement (µL level)
- High-throughput compatible (96/384-well plate format)
- Fast experiments (typically < 1 hour)
- Can be combined with CETSA for "in vitro + in vivo" dual validation