FPLC coupled with multi-angle light scattering

Protein samples (typically 100 µl at a concentration of 50 µM pre-equilibrated with column buffer) was injected into an AKTA FPLC system with a Superose-12 10/300 GL column (GE Healthcare) using the column buffer of 50 mM Tris-HCl (pH 7.8), 1 mM DTT, 1 mM EDTA, and 100 mM NaCl. The chromatography system was coupled to a multi-angle light scattering system equipped with a 18 angles static light scattering detector (Dawn, Wyatt) and a differential refractive index detector (Optilab, Wyatt). The elution profiles were analyzed using the ASTRA 6 software (Wyatt).