SEC-MALS was performed in reaction buffer (20 mM Tris-HCl, pH 8.0, 120 mM KOAc, 5 mM Mg(OAc)2, 2 mM DTT). 1 mg/ml of ρ were mixed with 2 mg/ml of Rof and incubated at 32 °C for 10 min. 60 μl of the mixture were loaded on a Superose 200 increase 10/300 GL column (Cytiva) and chromatographed on HPLC system (Agilent) coupled to a miniDAWN TREOS multi-angle light scattering and a RefractoMax 520 detector system (Wyatt Technologies). Prior to measurements, a system calibration was performed using BSA (Sigma-Aldrich). ASTRA 6.1 software was used for data analysis and molecular mass determination.
Superose 200 increase 10 300 gl column
Manufactured by Cytiva
The Superose 200 increase 10/300 GL column is a size exclusion chromatography column designed for the separation and purification of biomolecules, such as proteins and peptides, based on their size and molecular weight. It features a dextran-based stationary phase and a packed bed volume of 24 ml, suitable for use in low-to-medium pressure liquid chromatography systems.
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2 protocols using superose 200 increase 10 300 gl column
1
SEC-MALS Analysis of ρ-Rof Complex
2
SEC-MALS Analysis of ρ-Rof Complex
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SEC-MALS was performed in reaction buffer (20 mM Tris-HCl, pH 8.0, 120 mM KOAc, 5 mM Mg(OAc)2, 2 mM DTT). 1 mg/ml of ρ were mixed with 2 mg/ml of Rof and incubated at 32 °C for 10 min. 60 µl of the mixture were loaded on a Superose 200 increase 10/300 GL column (Cytiva) and chromatographed on HPLC system (Agilent) coupled to a miniDAWN TREOS multi-angle light scattering and a RefractoMax 520 detector system (Wyatt Technologies). Prior to measurements, a system calibration was performed using BSA (Sigma-Aldrich). ASTRA 6.1 software was used for data analysis and molecular mass determination.
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