Wtc 015s5 sized exclusion column

The purified PML RING_1–119 and mutants were subjected to gel filtration analysis (S100 column, GE Healthcare). The elution peaks, as monitored by UV absorption at 280 nm, were pooled separately and chosen for size exclusion chromatography-multi-angle light scattering (SEC-MALS) characterization, respectively. In brief, the purified protein samples were concentrated and analyzed using a WTC-015S5 sized exclusion column (Wyatt Technology) which was connected to a 1260 infinity liquid chromatography system (Agilent Technology) equipped with inline DAWN HELEOS-II MALS and Optilab rEX differential refractive index detectors (Wyatt Technology). For each sample, a 40 μl injection volume and 0.5 ml min⁻¹ flow rate were applied. Data were recorded and processed using ASTRA VI software (Wyatt Technology).

SafDAA-dsc and mutants were subjected to gel filtration analysis using a WTC-015S5 sized exclusion column (Wyatt Technology). The elution of each sample was analyzed by a 1260 infinity liquid chromatography system (Agilent Technology) linked with inline DAWN HELEOS-II MALS and Optilab rEX differential refractive index detectors (Wyatt Technology). For each run, a 40-μl sample (2.5 mg/ml) was injected. The sample was eluted at a flow-rate of 0.5 ml/min for SafDAA-dsc and mutants. Data were recorded and analyzed using ASTRA VI software (Wyatt Technology).