T*

Stability/Folding

Temperature of Maximum Stability

°C (Celsius)

Differential Scanning Calorimetry (DSC)

50 mM potassium phosphate

7.5

100 μM

T* is temperature of maximum stability (where maximal ΔG value is reached). See ΔG(T*) assay for how ΔG is calculated.

ΔG(T*) assay:
ΔG(T*) is Gibbs free energy at maximum stability (T* is temperature at which maximal ΔG value is reached).

Calorimetric data was fit in two stages to obtain Tm, ΔHm, and ΔCp (see Tm assay for details). From this modeled data, other parameters were derived.
The modified Gibbs-Helmotz equation (Becktel and Schellman, 1987) was used to calculate the temperature profiles of ΔGunfolding for each variant:

ΔG(T) = ΔH(Tm) (1-T/Tm) + ΔCp [T - Tm - T In(T/Tm)]