In a temperature-jump apparatus, a solution containing the reactant(s) at equilibrium conditions is rapidly heated by a high-voltage discharge (10-12kV) through an observation cell. The rapid increase in temperature perturbs the system equilibrium and the relaxation to a new equilibrium is observed by recording a change of a spectroscopical signal (fluorescence) as a function of time. T-jump apparatus can resolve fast reaction kinetics occurring in the µs time range. The use of specific optical filters allows detection of different fluorescence emission wavelength ranges.
Generally suitable to monitor ultrafast kinetics of reactions involving protein-protein interactions, protein (un)folding, enzyme activity, ligand binding, Forster resonance energy transfer (FRET).
Sample requirements:
- Pure protein samples are strongly suggested.
- Protein concentrations may vary for different types of experiments and should be previously discussed with the TNA site.
What other specific considerations are relevant?
A wide range of buffers can be used, however buffers with at least 150 mM of ionic strength are required to ensure conductivity of the solution.
Partners offering this technique
MOSBRI reference partner site for this technique: