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Heleos 2 multiangle

Manufactured by Wyatt Technology

The HELEOS-II multiangle is a light scattering instrument designed for the characterization of macromolecules and nanoparticles in solution. It measures the angular dependence of the scattered light, providing information on the size, shape, and interactions of the analytes.

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3 protocols using heleos 2 multiangle

1

Multiangle Light Scattering Analysis of Protein Samples

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Experiments were conducted on a system comprising a Wyatt HELEOS-II multiangle light scattering detector and a Wyatt rEX refractive index detector linked to a Shimadzu HPLC system (SPD-20A UV detector, LC20-AD isocratic pump system, DGU-20A3 degasser and SIL-20A autosampler). Work was conducted at room temperature (20 ± 2 °C). Sample injection volume was 100 μl at a protein concentration of 5 mg/ml. The samples were separated on a Superdex S200 10/300 (GE Healthcare) using 80 mm MES (pH 5.5), 200 mm NaCl as buffer. Shimadzu LC Solutions software was used to control the HPLC and Astra V software for the HELEOS-II and rEX detectors. Data were analyzed using the Astra V software. Molecular weights were estimated using the Zimm fit method (33 (link)) with 1 degree. A value of 0.19 was used for protein refractive index increment (dn/dc).
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2

Multiangle Light Scattering Protein Analysis

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Experiments were conducted on a system comprising a Wyatt HELEOS-II multiangle light scattering detector and a Wyatt rEX refractive index detector, linked to a Shimadzu HPLC system (SPD-20A UV detector, LC20-AD isocratic pump system, DGU-20A3 degasser and SIL-20A autosampler). Work was conducted at room temperature (20 ± 2°C). Sample injection volume was 100 μL at a protein concentration of 0.8 mg/mL. The samples were separated on a Superdex S200 10/300 (GE Healthcare) using 20 mM HEPES (pH 7.4), 200 mM NaCl as buffer. Shimadzu LC Solutions software was used to control the HPLC and Astra V software for the HELEOS-II and rEX detectors. Data were analyzed using the Astra V software. Molecular weights were estimated using the Zimm fit method (Zimm, 1945 ) with 1 degree. A value of 0.182 was used for protein refractive index increment (dn/dc).
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3

SEC-MALLS Analysis of OGT Protein

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SEC-MALLS analysis was conducted on a system comprising a Wyatt HELEOS-II multi-angle light scattering detector and a Wyatt rEX refractive index detector linked to a Shimadzu HPLC system (SPD-20A UV detector, LC20-AD isocratic pump system, DGU-20A3 degasser and SIL-20A autosampler). Experiments were conducted at room temperature. A Superdex S200 10/300 GL column pre-equilibrated in running buffer (20 mM HEPES 7.5, 200 mM NaCl) was used for separation. Sample injection was 100 µl of 3 mg/ml OGT. Flow rate was set at 0.5 ml/min. Shimadzu LabSolutions software was used to control the HPLC and Astra 7 software for the HELEOS-II and rEX detectors. The Astra data collection was 1 min shorter than the LC solutions run to maintain synchronisation. Data were analysed using the Astra 7 software and figures created using GraphPad Prism 5. MWs were estimated using the Zimm fit method with degree 1. A value of 0.182 was used for protein refractive index increment (dn/dc).
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