Minidawn treos 3 angle static light scattering detector

The sample of purified rSTEAP1 (50 μL) was injected onto a Shodex KW803 analytical SEC column mounted on a Waters HPLC system (Milford, MA) and eluted with the buffer containing 0.05% MNG-DDM at a rate of 0.5 mL/min. The mass of the eluted rSTEAP1 was measured using light scattering signals recorded with a Wyatt miniDAWN TREOS 3 angle-static light-scattering detector (Santa Barbara, CA), a Wyatt Optilab rEX refractive index detector and a Waters 2489 UV absorbance detector (21 (link)). The differential refractive index (dn/dc) for MNG-DDM, 0.128 mL/g, was calculated using the Wyatt refractive index detector. The size of the protein-detergent conjugate was de-convoluted following the published method (22 (link)). In these calculations, contributions from any co-purifying lipids were not distinguished from those of MNG-DDM.

The molecular mass of SEC-purified NaCT was measured using multi-angle light scattering ^{47 ,48} . DDM-purified NaCT sample (50 μL) was injected onto a Shodex KW803 analytical SEC column on a Waters HPLC (Milford, MA) and eluted with the buffer containing 0.05% DDM at a rate of 0.5 mL/min. The mass of the NaCT protein was determined using a Wyatt miniDAWN TREOS 3 angle-static light scattering detector (Santa Barbara, CA), a Wyatt Optilab rEX refractive index detector and a Waters 2489 UV absorbance detector. The differential refractive index (dn/dc) for DDM, 0.128 mL/g, was calculated using the refractive index detector. The size of the protein–detergent conjugate was deconvoluted following the published method ⁴⁹ , in which contributions from co-purifying lipids were not distinguished from those of the detergent.