SEC-MALS was performed using an ÄKTApure system (GE Healthcare) coupled to a miniDAWN TREOS II detector and an OptiLab T-rEX online refractive index detector (Wyatt Technology). The absolute molar mass was calculated by analysing the scattering data with the ASTRA analysis software package, version 7.2.2.10 (Wyatt Technology). Bovine serum albumin (BSA) was used for calibration, and proteins were separated on a Superdex 75 10/300 analytical column (GE Healthcare) at a flow rate of 0.4 ml/min. CodY (200 μl of 35 μM) was injected and eluted in 20 mM Tris–HCl pH 8, 200 mM NaCl. The refractive index increment was set at 17.66 μM for EfCodY and 5.37 μM for SaCodY, and the extinction coefficient for ultraviolet detection at 280 nm was calculated from the primary sequences.
Minidawn treos 2 detector
Manufactured by Wyatt Technology
The MiniDAWN TREOS II is a multi-angle light scattering (MALS) detector designed for use in size-exclusion chromatography (SEC) and other liquid chromatography systems. The device measures the angular dependence of scattered light intensity to determine the absolute molar mass and size of macromolecules and nanoparticles in solution.
Automatically generated - may contain errors
Lab products found in correlation
Kta pure system, by GE Healthcare (2 mentions)
Astra analysis software package, by Wyatt Technology (2 mentions)
Rid 20a, by Shimadzu (2 mentions)
Astra 7, by Wyatt Technology (2 mentions)
Spd m30a, by Shimadzu (2 mentions)
Superdex 75 10 300 gl, by GE Healthcare (2 mentions)
Prominence hplc system, by Shimadzu (2 mentions)
Optilab t rex online, by Wyatt Technology (1 mentions)
Superdex 75 10 300 analytical column, by GE Healthcare (1 mentions)
Superdex 200 10 300 analytical column, by GE Healthcare (1 mentions)
Optilab t rex, by Wyatt Technology (1 mentions)
4 protocols using minidawn treos 2 detector
1
SEC-MALS Analysis of CodY Proteins
2
Determining Odin Protein's Molar Mass
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OdinAK elution peak was further analyzed by SEC–multiangle light scattering (MALS) using an ÄKTApure system (GE Healthcare) coupled to a miniDAWN TREOS II detector and an OptiLab T-rEX online refractive index detector (Wyatt Technology). The absolute molar mass was calculated by analyzing the scattering data using ASTRA analysis software package, version 7.2.2.10 (Wyatt Technology). Bovine serum albumin (BSA) was used for calibration, and proteins were separated on a Superdex 200 10/300 analytical column (GE Healthcare) with a flow rate of 0.4 ml/min. Sample (200 μl) at 2 mg/ml was injected and eluted in 30 mM Mops (pH 7) and 150 mM NaCl buffer. The refractive index increment of Odin was set at 0.185 ml/g, and the extinction coefficient for ultraviolet detection at 280 nm was calculated from the primary sequence. The calculated molecular weight is the average from three runs, with the error being the SD.
3
Protein Characterization by MALS
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Protein molecular mass and heterogeneity were determined by multi-angle light scattering (MALS) using a miniDAWN TREOS II detector (Wyatt Technologies), coupled to a Shimadzu Prominence HPLC system with RID-20A (RI) and SPD-M30A (diode array) detectors. SEC to separate oligomeric species was performed using Superdex 75 10/300 GL or Superdex 200 15/150 GL increase columns (GE Healthcare) in SEC buffer. The protein concentration was 1–10 mg/ml, and the injected protein amount 15–150 µg. Data processing, baseline reduction, and molecular weight calculation were done in ASTRA 7 (Wyatt Technologies).
4
Protein Molecular Mass Analysis by MALS
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Protein molecular mass and heterogeneity were determined by multi-angle light scattering (MALS) using a miniDAWN TREOS II detector (Wyatt Technologies), coupled to a Shimadzu Prominence HPLC system with RID-20A (RI) and SPD-M30A (diode array) detectors. SEC to separate oligomeric species was performed using Superdex 75 10/300GL or Superdex 200 15/150GL increase columns (GE Healthcare) in SEC buffer. The protein concentration was 1-10 mg/ml, and the injected protein amount 15-150 µg. Data processing, baseline reduction, and molecular weight calculation were done in ASTRA 7 (Wyatt Technologies).
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