Molecular masses of PLP and DM20 detergent complexes were determined using SEC-MALS at the core facility for Biophysics, Structural Biology, and Screening (BiSS) at the University of Bergen. A Superdex 200 Increase 10/300 (GE Healthcare) column was used to separate empty detergent micelles from the protein-detergent complexes. 50 µg of PLP or DM20 in SEC buffer (0.2% DM, 0.3 M NaCl, 0.5 mM TCEP, 5% glycerol, 20 mM Tris pH 7.5) was injected into the SEC column using Shimadzu Prominence I LC-2030C HPLC unit, and UV absorption was recorded by LC-2030/2040 PDA detector (Shimadzu). The light scattering was measured by a Wyatt miniDAWN TREOS instrument (Wyatt Technology) and refractive index was determined using a RefractoMax 520 detector (ThermoScientific). Data were analysed using Astra 7 (Wyatt Technology) utilizing protein conjugate analysis with dn/dc values of 0.1473 ml/g and 0.1850 ml/g for DM and proteins, respectively.
Prominence 1 lc 2030c hplc unit
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The Prominence I LC-2030C HPLC unit is a high-performance liquid chromatography system designed for analytical and preparative applications. It features a compact and integrated design, providing a complete HPLC solution in a single unit. The core function of the Prominence I LC-2030C is to perform liquid chromatographic separations and analyses of a wide range of chemical compounds.
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2 protocols using prominence 1 lc 2030c hplc unit
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SEC-MALS Characterization of PLP and DM20
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SEC-MALS Analysis of PLP and DM20
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Molecular masses of PLP and DM20 detergent complexes were determined using SEC-MALS at the core facility for Biophysics, Structural Biology, and Screening (BiSS) at the University of Bergen. A Superdex 200 Increase 10/300 (GE Healthcare) column was used to separate empty detergent micelles from the protein-detergent complexes. 50 µg of PLP or DM20 in SEC buffer (0.2% DM, 0.3 M NaCl, 0.5 mM TCEP, 5% glycerol, 20 mM Tris pH 7.5) was injected into the SEC column using Shimadzu Prominence I LC-2030C HPLC unit, and UV absorption was recorded by LC-2030/2040 PDA detector (Shimadzu). The light scattering was measured by a Wyatt miniDAWN TREOS instrument (Wyatt Technology) and refractive index was determined using a RefractoMax 520 detector (ThermoScientific). Data were analysed using Astra 7 (Wyatt Technology) utilizing protein conjugate analysis with dn/dc values of 0.1473 ml/g and 0.1850 ml/g for DM and proteins, respectively.
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