Name:

Tm

Category:

Stability/Folding

Property:

Melting Temperature (Tm)

Units:

°C (Celsius)

Measurement Technique:

Circular Dichroism (CD) , Thermal Denaturation

Buffers:

None

Temperature:

20-100 °C with 1 °C/min increments

pH:

None

Protein Concentration:

25-50 μM

Wavelength:

222 nm

Ionic Strength:

None

Details:

Thermal denaturation curves were obtained in SB250buffer using 25-50μM Ni-affinity purified protein in a 2.0 mm path-length cuvette. Samples were heated and cooled between 20°C and100C with a 2°C step size and 2 min sample equilibration time ateach temperature. At each temperature, ellipticity was monitored at222 nm using 60 s total data collection time, and 50 s integration time. Thermal melt fitting was performed as described assuming atwo-state model and a ΔCp value of 896 cal/mol K was set for all variants (14 cal/mol K per residue).