Asl fragments were individually injected (50 µM; 100 µl) onto a Superdex 200 10/300 GL size exclusion column (GE Healthcare) at 0.5 ml/min in buffer D (25 mM, Hepes, pH 7.5, 300 mM NaCl, 0.1% β-mercaptoethanol, and 0.2 g/liter sodium azide) and then passed consecutively through a UV detector, a light scattering instrument (DAWN HELEOS II; Wyatt Technology), and a refractometer (Optilab rEX; Wyatt Technology). The light scattering and refractive index data were used to calculate the weight-averaged molar mass using Wyatt Astra V software (Wyatt Technology). Data were processed with ASTRA software and plotted using SigmaPlot v11.
Wyatt astra 5 software
Manufactured by Wyatt Technology
The Wyatt Astra V software is a comprehensive data analysis tool designed for multi-angle light scattering (MALS) and size-exclusion chromatography (SEC) experiments. It provides advanced algorithms and algorithms for the determination of molecular weight, size, and other properties of macromolecules and nanoparticles.
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Lab products found in correlation
Dawn heleos 2, by Wyatt Technology (1 mentions)
Superdex 200 10 300 gl size exclusion column, by GE Healthcare (1 mentions)
Optilab rex, by Wyatt Technology (1 mentions)
Superdex 200 10 30 gl size exclusion column, by GE Healthcare (1 mentions)
Dawn heleos 2 multiangle static light scattering mals detector, by Wyatt Technology (1 mentions)
2 protocols using wyatt astra 5 software
1
Asl Fragment Size Exclusion Analysis
2
Size Exclusion Chromatography for Protein Analysis
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Size exclusion chromatography–multiangle light scattering (SEC-MALS) was performed as previously described (Klebba et al., 2015b ). Briefly, Asl mutants (50 μM, 100 μl) were injected onto a Superdex 200 10/30 GL size exclusion column (GE Healthcare Life Sciences) at 0.5 ml/min in buffer (25 mM HEPES, pH 7.5, 300 mM NaCl, 0.1% β-mercaptoethanol, and 0.2 g/l sodium azide). Eluent was analyzed by a tandem RI detector/Dawn Heleos II multiangle static light scattering (MALS) detector (Wyatt Technology). Light scattering and refractive index data were used to calculate the weight-averaged molar mass using Wyatt Astra V software (Wyatt Technology).
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