Following the method described by Yang et al. (15 (link)), the molecular weight of Qingke β-glucan was measured using SEC-MALLS-RI. By a DAWN HELEOS-II laser photometer (Wyatt, USA) equipped with an SB-803 HQ and an SB-804 HQ column (Showa Denko, Japan), the molecular weight of Qingkes β-glucans was measured in a 0.1 mol/L NaNO3 aqueous solution, including 0.03% NaNO3, with a flow rate of 0.45 ml/min. The temperature was monitored at 45°C by a model column heater (Sanshu Biotech, Shanghai, China). The concentration of the Qingke β-glucans is 1 mg/ml. A differential refractive index detector (Wyatt, USA) was used to detect the dn/dc value. The refractive index increment (dn/dc) value of the fractions was set to be 0.142 ml/g. Chromatographic data was processed with software ASTRA 6.1.
Differential refractive index detector
Manufactured by Wyatt Technology
Sourced in United States
The Differential Refractive Index Detector is a laboratory instrument used to measure the refractive index difference between two fluid streams. It is designed to provide accurate and reliable data for a variety of analytical applications.
Automatically generated - may contain errors
Lab products found in correlation
Superdex 200, by GE Healthcare (3 mentions)
Astra software package, by Wyatt Technology (3 mentions)
Astra software, by Wyatt Technology (2 mentions)
Superdex 200 10 300 gl, by GE Healthcare (2 mentions)
Sb 803 hq, by Resonac (1 mentions)
Dawn heleos 2 laser photometer, by Wyatt Technology (1 mentions)
Superdex 200 5 150 column, by Cytiva (1 mentions)
Hplc system, by Shimadzu (1 mentions)
Kta pure system, by GE Healthcare (1 mentions)
Superdex 200 increase 10 300 gl column, by GE Healthcare (1 mentions)
Superdex 200 increase 10 300 gl, by Cytiva (1 mentions)
Ft ir 6300, by Jasco (1 mentions)
515 hplc, by Waters Corporation (1 mentions)
12 protocols using differential refractive index detector
1
Qingke β-Glucan Molecular Weight Determination
2
Type ISP Enzymes Size Exclusion Chromatography
3
Purification and Characterization of SauUSI
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1 mg/ml of the SauUSI was injected into a GE Superdex 200™ column equilibrated with 50 mM Tris–HCl, pH 8, 50 mM NaCl and 1 mM DTT. The apparatus was connected to a light scattering diode array and a differential refractive index detector (Wyatt Technology). The data was analyzed using the ASTRA software 6.1.7.17. The molar mass was calculated from the light scatter and differential refractive index. Before use, the column was calibrated using bovine serum albumin.
4
Analytical Gel Filtration Chromatography
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100 μL Rpd3S sample (40 μM) was loaded onto a Superdex 200 5/150 column (Cytiva) with a buffer containing 20 mM Tris-HCl pH 7.5, 150 mM NaCl. A static light-scattering detector and a differential refractive index detector (Wyatt) were connected to the analytical gel filtration chromatography system. Data were analyzed with ASTRA7 provided by Wyatt.
5
Type ISP Enzymes Size Exclusion Chromatography
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The Type ISP enzymes (0.5-1.0 mg/ml) were run in reaction buffer (for LlaGI, 50 mM Tris-Cl, pH 8.0, 10 mM MgCl2, 1 mM DTT; for LlaBIII, 50 mM Tris-Cl, pH 8.0, 10 mM MgCl2, 150 mM KCl, 1 mM DTT) at 1 ml/min on a Superdex 200 analytical size exclusion column (GE Healthcare), attached to a light scattering diode array and a differential refractive index detector (Wyatt Technology). Chromatograms were analysed using the ASTRA software (v6.0.5.3, Wyatt Technology). Absolute molecular masses of elutes were calculated from the ratio of light scattering and the differential refractive index. Reproducibility of the column chromatography was checked using bovine serum albumin as in Butterer et al.55
6
Molecular Mass Determination by SEC-MALS
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Experiments were performed on an HPLC system (Shimadzu) equipped with autosampler and UV detector, using a preequilibrated analytical SEC column (Superdex 200 10/300 GL; GE Healthcare) with gel filtration buffer. A sample of the purified CNBHD (2 mg/ml, 50 µl) was loaded onto the column and analyzed using an inline eight-angle light-scattering detector, followed by a differential refractive-index detector (Wyatt Technology). Refractive index and MALS readings were analyzed using the Astra software package (Wyatt Technology) to determine the molecular mass.
7
Analytical Gel Filtration Chromatography
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Analytical gel filtration chromatography was carried out on an ÄKTA Pure system (GE Healthcare). Protein samples at indicated concentration were loaded onto a Superdex 200 Increase 10/300 GL column (GE Healthcare) equilibrated with 50 mM Tris-HCl buffer, pH 7.5, containing 100 mM NaCl.
The static light-scattering detector and differential refractive index detector (Wyatt) were coupled to the analytical gel filtration chromatography system. Data were analyzed with ASTRA6 provided by Wyatt.
The static light-scattering detector and differential refractive index detector (Wyatt) were coupled to the analytical gel filtration chromatography system. Data were analyzed with ASTRA6 provided by Wyatt.
8
Molecular Mass Determination of mTTYH1-GFP-8xHis
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Experiments were performed using an analytical SEC column (Superdex 200 Increase 10/300 GL; Cytiva) preequilibrated with gel filtration buffer (20 mM Tris, pH 7.5, 150 mM NaCl, 0.3 mM DDM). Samples of purified mTTYH1-GFP-8xHis (50 μL, 1.18 mg/mL) were loaded onto the SEC column connected in line to a downstream 8-angle light scattering detector, followed by a differential refractive index detector (Wyatt Technology). Protein conjugate analysis was performed using a dedicated program within the Astra software package (Wyatt Technology) to determine the molecular mass of the protein and detergent components of the complex.
9
Polymer Molecular Weight Analysis
10
Size-Exclusion Chromatography for Molecular Mass
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Experiments were carried out using a size-exclusion chromatography column (superdex-200 increase 10/300 GL column) pre-equilibrated with Tris-HCl, pH 7.5, 150 mM NaCl, 20 mM β-mercaptoethanol, and 0.02% Triton X-100. Samples (2 mg/mL; 50 µL) were injected onto an HPLC, connected to an eight-angle light-scattering detector, followed by a differential refractive-index (RI) detector (Wyatt Technology, Santa Barbara, CA, USA). RI and MALS readings were analyzed with the ASTRA software package (Wyatt Technology, Santa Barbara, CA, USA) to determine molecular mass.
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