The purity, molecular mass and size distribution of the proteins were analyzed by an analytical light scattering instrument (SEC-MALS) consisting of an Agilent 1260 Infinity Isocratic Liquid Chromatography System, a Wyatt Dawn Heleos II Multi-Angle Light Scattering Detector (Wyatt Technology) and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). Analytical size exclusion chromatography was performed at room temperature using a Superdex 200 10/300 GL column (GE Healthcare) equilibrated with a mobile phase containing 10 mM HEPES (pH 7.4), 200 mM NaCl, and 5 mM β-ME. 100 μl protein sample at concentration of 2 mg/ml or 12 mg/ml was injected into the column and eluted at a flow rate of 0.4 ml/min. The column effluent was monitored in-line with three detectors that simultaneously monitor the UV absorption, light scattering and refractive index. The measurements were analyzed using the ASTRA software (Wyatt Technology) to determine the molecular mass of the protein42 (link).
Wyatt optilab t rex refractive index detector
Manufactured by Wyatt Technology
Sourced in United States
The Wyatt Optilab T-rEX Refractive Index Detector is a laboratory instrument used to measure the refractive index of a sample. It is designed to provide accurate and reliable measurements of the refractive index of liquids, solutions, and other materials. The T-rEX detector incorporates a laser-based optical system and temperature control to ensure precise and consistent measurements.
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Lab products found in correlation
Wyatt dawn heleos 2 multi angle light scattering detector, by Wyatt Technology (9 mentions)
Superdex 200 10 300 gl column, by GE Healthcare (7 mentions)
Astra software, by Wyatt Technology (5 mentions)
Astra 7, by Wyatt Technology (2 mentions)
Astra 6, by Wyatt Technology (1 mentions)
Wtc 030s5 column, by Wyatt Technology (1 mentions)
Bovine serum albumin (bsa), by Thermo Fisher Scientific (1 mentions)
Agilent 1260 infinity 2 liquid chromatography system, by Agilent Technologies (1 mentions)
10 protocols using wyatt optilab t rex refractive index detector
1
Protein Characterization by SEC-MALS
2
Protein Purity and Molecular Weight Analysis
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The purities and molecular weights of the proteins were analyzed by an analytical light scattering instrument (SEC-MALS) consisting of an Agilent 1260 Infinity Isocratic Liquid Chromatography System, a Wyatt Dawn Heleos II Multi-Angle Light Scattering Detector, and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). Analytical size exclusion chromatography was performed at 24 °C using a Superdex 200 10/300 GL column (GE Healthcare) equilibrated with a mobile phase containing 10 mM HEPES (pH 7.4), 200 mM NaCl, and 5 mM β-ME. Hundred microliter protein solution was injected into the column and eluted at a flow rate of 0.4 ml/min. The column effluent was monitored simultaneously with three detectors for UV absorption, light scattering and refractive index. The data were analyzed using the ASTRA software (Wyatt Technology) to determine the molecular mass of the protein44 (link).
3
Protein Characterization by SEC-MALS
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The purity, molecular mass, and size distribution of the proteins were analyzed by an analytical light scattering instrument (SEC-MALS) consisting of an Agilent 1260 Infinity Isocratic Liquid Chromatography System, a Wyatt Dawn Heleos II multiangle light scattering detector (Wyatt Technology), and a Wyatt Optilab T-rEX refractive index detector (Wyatt Technology). Analytical SEC was performed at about 25°C using a Superdex 200 10/300 GL column (GE Healthcare) equilibrated with a mobile phase containing 10 mM Hepes (pH 7.5), 200 mM NaCl, and 10 mM DTT. One hundred microliters of protein sample (10 mg/ml) was injected into the column and eluted at a flow rate of 0.5 ml/min. The column effluent was monitored in-line with three detectors that simultaneously monitor ultraviolet absorption, light scattering, and the refractive index. The measurements were analyzed using the ASTRA software (Wyatt Technology) to determine the molecular mass of the protein. The experiments were performed three times to check for reproducibility of the results.
4
Protein Characterization by SEC-MALS
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Purity, molecular mass and size distribution of the purified proteins were analyzed using an analytical light scattering instrument (SEC-MALS) equipped with an Agilent 1260 Infinity Isocratic Liquid Chromatography System, a Wyatt Dawn Heleos II Multi-Angle Light Scattering Detector (Wyatt Technology), and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). Analytical size exclusion chromatography was performed at room temperature using a Superdex 200 10/300 GL column (GE Healthcare) equilibrated with a buffer containing 10 mM HEPES (pH 7.5) and 100 mM NaCl. 50 μl protein sample at concentration of about 1.5 mg ml−1 was injected with a flow rate of 0.5 ml min−1. The column effluent was monitored in-line with three detectors that simultaneously monitor the UV absorption, light scattering, and refractive index. The measurements were analyzed using the ASTRA 6.1 software (Wyatt Technology) to determine the molecular masses of the proteins.
5
SEC-MALS Analysis of Protein Purity
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The purities and molecular weights of the proteins were analyzed by a SEC-MALS instrument consisting of an Agilent 1260 Infinity Isocratic Liquid Chromatography System, a Wyatt Dawn Heleos II Multi-Angle Light Scattering Detector, and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). Analytical SEC was performed at 24 °C using a Superdex 200 10/300 Gl column (GE Healthcare) equilibrated with a mobile phase containing 10 mM Hepes (pH 7.4), 200 mM NaCl, and 5 mM β-ME. Hundred microliters protein solution was injected into the column and eluted at a flow rate of 0.4 ml/min. The column effluent was monitored simultaneously with three detectors for UV absorption, light scattering, and refractive index. Light scattering detector is used for molecular weight measurement; UV and refractive index detectors are used for concentration measurements in two orthogonal ways. The data were analyzed using the ASTRA software (Wyatt Technology) to determine the molecular weight of the protein (29 (link)).
6
SEC-MALS Analysis of cMCU-ΔNTD Protein
7
SEC-MALS Analysis of cMCU-ΔNTD Protein
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The instrument setup used for the SEC-MALS experiment consisted of an Agilent 1260 Infinity Isocratic Liquid Chromatography System connected in series with a Wyatt Dawn Heleos II Multi-Angle Light Scattering (MALS) detector (Wyatt Technology) and a Wyatt Optilab T-rex Refractive Index Detector (Wyatt Technology). Analytical size-exclusion chromatography was performed at room temperature using Superdex 200 10/300 GL column (GE Healthcare) equilibrated with a mobile phase containing 20 mM MES (pH 6.4), 75 mM NaCl, 0.48 mM Foscholine-14, 0.3 mM NaN3, and 2 mM EDTA. 100 μl purified cMCU-ΔNTD sample at 4.0 mg/ml was injected into the column and eluted at a flow rate of 0.4 ml/min. The column effluent was monitored in-line with three detectors that simultaneously monitored UV absorption, light scattering (LS), and refractive index (RI), respectively. The data from the three detectors were imported by the ASTRA software package, and the three-detector method28 was used to determine the molecular mass.
8
Static Light Scattering of Proteins
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The static light scattering (SLS) experiments were performed using an Agilent 1260 Infinity Isocratic Liquid Chromatography System coupled with a Wyatt Dawn Heleos II Multi-Angle Light Scattering (MALS) detector (Wyatt Technology) and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). 40 μL samples were loaded onto the WTC-030S5 column (Wyatt Technology) with a mobile phase containing 150 mM NaCl, 20 mM HEPES, pH 7.5 at a flow rate of 0.5 mL min−1. BSA (ThermoFisher Scientific) was used to calibrate the static light scattering system. Data were recorded and processed using Astra 7 software (Wyatt Technology).
9
Molecular Weight Analysis of Polyelectrolyte Complexes
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The molecular weight (MW) distribution of PEC, PPH and POS were analyzed by injecting 100 μL of 2.5 mg/mL substrate to Agilent 1,260 Infinity II liquid chromatography system (Agilent, United States) equipped with PSS precolumn 10 μm 8 × 50 mm, PSS Suprema Lux analytical Ultrahigh, and PSS Suprema Lux 30 Å (PSS, Germany). The separation was performed using 0.3 M lithium acetate buffer as eluent at flow rate 1.0 mL/min, and column temperature was 37°C. Wyatt Dawn 8+ light scattering instrument and Wyatt Optilab T-rex refractive index detector (Wyatt, United States) were used for detection. MW and concentrations were converted from the detector signals by the software ASTRA 7 (Wyatt).
10
Protein Molar Mass Analysis by SEC-MALS
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The molar mass of the purified protein was analyzed by SEC–MALS on an Agilent 1260 Infinity Isocratic Liquid Chromatography System (Agilent) incorporated with a Wyatt Dawn Heleos II Multi-Angle Light Scattering Detector and a Wyatt Optilab T-rEX Refractive Index Detector (Wyatt Technology). Protein solution (2 mg/ml) was injected into a Superdex 200 10/300 column with a mobile phase containing 20 mM Hepes (pH 7.4) and 200 mM NaCl at a flow rate of 0.4 ml/min. The eluate was monitored with three detectors for UV absorption, light scattering, and refractive index. The UV and refractive index detectors were used to quantify the protein concentration in two orthologous ways; and the light scattering detector was used to determine the protein molar mass. The data were analyzed using the ASTRA software (Wyatt Technology) to determine the molar mass of the protein.
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