Size exclusion chromatography coupled with multiangle
light scattering (SEC-MALS) analysis was performed at 25 °C.
A 500 μL portion of 1.5 mg/mL protein was loaded onto a size
exclusion column (for the native protein a Superose 6 10/300 GL column
was used and for the dimer mutant a Superdex 200 10/300GL column (GE
Life Sciences)), equilibrated in 200 mM KCl, 1 mM MnCl2, 50 mM Tris pH 7.5 and a flow rate of 0.75 mL/min. Eluting samples
were passed through a Wyatt DAWN Heleos II EOS 18-angle laser photometer
coupled to a Wyatt Optilab rEX refractive index detector. Data were
analyzed using Astra 6 software (Wyatt Technology Corp., CA, USA).
light scattering (SEC-MALS) analysis was performed at 25 °C.
A 500 μL portion of 1.5 mg/mL protein was loaded onto a size
exclusion column (for the native protein a Superose 6 10/300 GL column
was used and for the dimer mutant a Superdex 200 10/300GL column (GE
Life Sciences)), equilibrated in 200 mM KCl, 1 mM MnCl2, 50 mM Tris pH 7.5 and a flow rate of 0.75 mL/min. Eluting samples
were passed through a Wyatt DAWN Heleos II EOS 18-angle laser photometer
coupled to a Wyatt Optilab rEX refractive index detector. Data were
analyzed using Astra 6 software (Wyatt Technology Corp., CA, USA).