The dissolved sunflower seed protein and hydrolyzed sunflower seed protein were filtered through a 0.22 μm filter. Then, the samples were analyzed with a HELEOS system gel permeation chromatography (Wyatt, America) coupled with the Wyatt DAWN HELEOS-II laser detector and the Wyatt Optilab-rEX differential detector equipped with a Shodex OH-pak SB-806 column. The working conditions include ultra-pure water (0.2 mol L−1 potassium dihydrogen phosphate and 0.02% sodium azide) as the mobile phase, 0.5 mL min−1 of flow rate, 35 °C of column temperature and 500 μL of infusion volume.
Dawn heleos 2 laser detector
Manufactured by Wyatt Technology
The DAWN HELEOS-II is a laser detector manufactured by Wyatt Technology. It is designed to measure the light scattering properties of samples in a laboratory setting. The device uses a laser to illuminate the sample and detects the scattered light at multiple angles, providing information about the size, shape, and concentration of particles within the sample.
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2 protocols using dawn heleos 2 laser detector
1
Sunflower Seed Protein Characterization
2
Molecular Weight Determination of COMsrA
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SEC coupled to multiangle laser light scattering (SEC-MALLS) was used to determine the solution molecular weight of full-length and ΔH-domain forms of CoMsrA. Protein samples at concentrations of 2.5 and 1.5 mg/ml for full-length and ΔH-domain forms, respectively, were applied to a WTC-015S5 column mounted on a Shimadzu HPLC system equilibrated at a flow rate of 0.5 ml/min with 20 mM Tris-HCl (pH 8.0) and 100 mM NaCl at 298K. The scattered light intensity and protein concentration of the column eluate were measured using a DAWN HELEOS II laser detector and an OPTILAB T-rEX refractive index detector (Wyatt Technology), respectively. Data analysis from both detectors was carried by ASTRA software version 6.1 (Wyatt Technology).
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