100 μl of Ctf4CTD protein at a concentration of 2 mg/ml was loaded onto a Superdex S200 HR 10/300 gel-filtration column (GE Healthcare) in 25 mM Hepes pH 7.0, 200 mM NaCl at a flow rate of 0.5 ml/min. The column was controlled using an Äkta Purifier System (GE Healthcare) and was linked to a DAWN 8+ 8-angle light scattering detector (Wyatt Technology) with a fused silica sample cell using a laser wavelength of 664 nm. The change in the refractive index was detected using an Optilab T-rEX refractometer with extended range (Wyatt Technology) using a wavelength of 658 nm. Data collection and analysis was carried out using the ASTRA6 software package (Wyatt Technology). Molecular weight determination across the sample peak was carried out using a Zimm-plot derived global fitting algorithm with a fit degree of 1 and a dn/dc value of 0.1850 ml/g.
Superdex s200 hr 10 300
Manufactured by GE Healthcare
The Superdex S200 HR 10/300 is a size-exclusion chromatography column used for the separation and purification of proteins, peptides, and other biomolecules. The column features a high-resolution bed material that allows for efficient separation of molecules based on their size and molecular weight.
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2 protocols using superdex s200 hr 10 300
1
Ctf4 Protein Characterization by SEC-MALS
2
Ctf4 Protein Characterization by SEC-MALS
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100 μl of Ctf4CTD protein at a concentration of 2 mg/ml was loaded onto a Superdex S200 HR 10/300 gel-filtration column (GE Healthcare) in 25 mM Hepes pH 7.0, 200 mM NaCl at a flow rate of 0.5 ml/min. The column was controlled using an Äkta Purifier System (GE Healthcare) and was linked to a DAWN 8+ 8-angle light scattering detector (Wyatt Technology) with a fused silica sample cell using a laser wavelength of 664 nm. The change in the refractive index was detected using an Optilab T-rEX refractometer with extended range (Wyatt Technology) using a wavelength of 658 nm. Data collection and analysis was carried out using the ASTRA6 software package (Wyatt Technology). Molecular weight determination across the sample peak was carried out using a Zimm-plot derived global fitting algorithm with a fit degree of 1 and a dn/dc value of 0.1850 ml/g.
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