The SEC-MALS analysis was performed on a DAWN HELEOS multi-angle light scattering detector, with eighteen angles detectors and a 658.9 nm laser beam, (Wyatt Technology, Santa Barbara, CA, USA) and an Optilab T-rEX refractometer (Wyatt Technology) in-line with two coupled Superdex 200 Increase 10/300 GL size exclusion chromatography analytical columns. Experiments were performed using an isocratic pump (Agilent) with a flow of 0.5 ml/min at room temperature (25°C). Data collection was performed with ASTRA 6.1 software (Wyatt Technology). For the experiments, 300 μl at 1.3 mg/ml protein were loaded on the columns with running buffer of 25 mM HEPES pH 8, 10% glycerol, 300 mM NaCl and 2 mM EDTA. Calibration was performed with high-molecular weight markers from GE Healthcare (thyroglobulin, 669 000 Da; ferritin, 440 000 Da; aldolase, 158 000 Da; conalbumin, 75 000 Da; and ovoalbumin, 43 000 Da).
Dawn heleos multi angle light scattering detector
Manufactured by Wyatt Technology
Sourced in United States
The DAWN HELEOS is a multi-angle light scattering (MALS) detector designed for the characterization of macromolecules and nanoparticles. The detector measures the intensity of light scattered at multiple angles, which can be used to determine the molar mass, size, and conformation of samples.
Automatically generated - may contain errors
Lab products found in correlation
Isocratic pump, by Agilent Technologies (1 mentions)
Thyroglobulin, by GE Healthcare (1 mentions)
Ferritin, by GE Healthcare (1 mentions)
Ovoalbumin, by GE Healthcare (1 mentions)
Aldolase, by GE Healthcare (1 mentions)
Conalbumin, by GE Healthcare (1 mentions)
658.9 nm laser beam, by Wyatt Technology (1 mentions)
Astra 6, by Wyatt Technology (1 mentions)
Optilab t rex refractometer, by Wyatt Technology (1 mentions)
Optilab rex refractometer, by Wyatt Technology (1 mentions)
Superose 6 10 300 gl column, by GE Healthcare (1 mentions)
2 protocols using dawn heleos multi angle light scattering detector
1
Protein Characterization by SEC-MALS
2
Molecular Weight Distribution Analysis of Polysaccharides
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The method of Shengping Xiang [32 (link)] was used to characterize the molecular weight distribution of PP from different origins. GPC-MALLS was used to characterize the PP from different areas as well as the PP adsorbed onto the emulsion interface and that present in aqueous phase. The system consists of a Superose 6 10/300GL column (GE Healthcare Co., Boston, MA, USA) and a DAWN HELEOS multi-angle light scattering detector (Wyatt Technology Corporation, Santa Barbara, CA, USA), an SPD-10Avp series UV detector (Shimadzu Technologies, Kyoto, Japan) operated at 214 nm, an Optilab rEX refractometer (Wyatt Technology Corporation, Santa Barbara, CA, USA) at 25 ± 0.2 °C. 0.2 M NaCl aqueous solution containing 0.03% NaN3 with a flow rate of 0.4 mL/min was used as an eluent after filtration through a 0.45 µm Millipore filter. A dn/dc value of 0.142 mL/g was taken for PP.
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