Melting Temperature (Tm)
30-90 °C with 2.0 °C/min increments
excitation at 266 nm and emission at 280-450 nm
Intrinsic protein fluorescence (266 nm excitation, 280-450 nm emission scan) and static light scattering (SLS) at 266 and 473 nm, were measured simultaneously with a UNit (Unchained Laboratories) at every 1 °C for 30–90 °C after 30-s equilibration at each temperature. Microcuvettes were loaded with 9 µL of 0.1 mg/mL samples in triplicate. Tagg was determined from SLS counts at 266 nm using the instrument software.