G. lucidum raw material (Ling-Zhi) was obtained from Wyntek Corporation (Taiwan). Two additional steps were required for the purification of the Ling-Zhi extract, namely the preparation of a crude extract and the preparation of the final Ling-Zhi extract. The compounds in the crude extract and in the Ling-Zhi extract were monitored by high-pressure liquid chromatography using a size exclusion column Tosoh TSKgel G5000PWXL. Detailed information about FFLZ and the preparations of the G. lucidum polysaccharide extract has been previously reported1 2 (link).
Tsk gel g5000pwxl
Manufactured by Tosoh
Sourced in Japan
The TSK-Gel G5000PWXL is a size exclusion chromatography (SEC) column designed for the analysis of high molecular weight polymers and proteins. It features a silica-based packing material with a pore size of 5,000 Angstroms, which allows for the separation of large macromolecules. The column is suitable for use in aqueous and organic mobile phases, making it a versatile tool for a variety of applications.
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Lab products found in correlation
G4000pwxl, by Tosoh (3 mentions)
Astra software, by Wyatt Technology (3 mentions)
Refractive index monitor, by Waters Corporation (2 mentions)
Dawn eos malls detector, by Wyatt Technology (2 mentions)
Astra 6, by Wyatt Technology (2 mentions)
High performance liquid chromatography (hplc), by Shimadzu (1 mentions)
2414 refractive index detector, by Waters Corporation (1 mentions)
G3000pwxl, by Tosoh (1 mentions)
Quasi elastic light scattering, by Wyatt Technology (1 mentions)
1260 infinity, by Agilent Technologies (1 mentions)
Tskgel g3000pwxl, by Tosoh (1 mentions)
Lc 6a, by Shimadzu (1 mentions)
10 protocols using tsk gel g5000pwxl
1
Purification of Ganoderma lucidum Extract
2
Alginate Molecular Weight Characterization
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Alginate molecular weight and polydispersity were characterized by size exclusion chromatography-multi-angle laser light scattering (SEC-MALLS) analysis14 (link). DAWN-EOS multi-angle laser light scattering detector with a laser at 690 nm (Wyatt Technology Corp., Santa Barbara, CA, USA); Waters 2410 refractive index monitor). Purified samples were dissolved in 0.1 M NaNO3 (2 mg/mL) and allowed to hydrate fully by incubating at room temperature overnight. Immediately prior to analysis, samples were pre-heated at 80 °C for 5 min, injected (100 μL) and eluted with 0.1 M NaNO3 (0.7 mL/min, 60 °C) from two columns (TSK-Gel G5000PWXL and G4000PWXL, 300 × 7.8 mm, Tosoh Corp.) connected in series. ASTRA software (version 6.1.2.84, Wyatt Technology Corp.) and dn/dc of 0.150 mL/g was used for determining weight-average molecular weights (Mr) and number-average molecular weights (Mn) and polydispersity index (PI) via the fraction Mw/Mn. In the case of a perfectly monodisperse (homogeneous) polymer PI value equals 1.0.
3
Protein Molecular Weight and Size Analysis
4
Molecular Weight Determination of ZOP
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The molecular weight of ZOP was determined by high performance gel permeation chromatography (HPGPC) (13 (link)). It was equipped with TSK-GEL G5000PWXL (300mm × 7.8mm, i.d.) and G3000PWXL (300mm × 7.8mm, i.d.) gel columns in series (Tosoh Biosep, Japan) and a Waters 2414 refractive index detector (Massachusetts, USA). The samples were eluted at 0.6 mL/min flow rate with monopotassium phosphate solution as mobile phase. The T-series dextran was used as standard.
5
Molecular Weight Analysis of Polymers
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Molar mass was determined using size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS). Samples (2 mg/mL in 0.1 mol L -1 NaNO3) were allowed to hydrate by standing at room temperature overnight and centrifuged (14,000 x g, 10 min) to clarify. The soluble material (100 µL) was injected onto two columns (TSK-Gel G5000PWXL and G4000PWXL, 300 x 7.8mm, Tosoh Corp., Tokyo, Japan) connected in series and eluted with 0.1 M NaNO3 (0.7 mL min -1 , 60 C). The eluted material was detected using a UV spectrophotometer (280 nm), a DAWN-EOS MALLS detector (Wyatt Technology Corp., Santa Barbara, USA) and a refractive index (RI) monitor (Waters Corp., Milford, USA). The data for molar mass determination was analysed using ASTRA software (v6.1.84, Wyatt Technology Corp.) using a refractive index increment, dn/dc, of 0.146 mL g -1 [24] .
6
Oligomerization of αA-crystallin Variants
7
Molecular Weight Analysis of Polymers
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Molar mass was determined using size-exclusion chromatography coupled with multi-angle laser light scattering (SEC-MALLS). Samples (2 mg/mL in 0.1 mol L -1 NaNO3) were allowed to hydrate by standing at room temperature overnight and centrifuged (14,000 x g, 10 min) to clarify. The soluble material (100 µL) was injected onto two columns (TSK-Gel G5000PWXL and G4000PWXL, 300 x 7.8mm, Tosoh Corp., Tokyo, Japan) connected in series and eluted with 0.1 M NaNO3 (0.7 mL min -1 , 60 C). The eluted material was detected using a UV spectrophotometer (280 nm), a DAWN-EOS MALLS detector (Wyatt Technology Corp., Santa Barbara, USA) and a refractive index (RI) monitor (Waters Corp., Milford, USA). The data for molar mass determination was analysed using ASTRA software (v6.1.84, Wyatt Technology Corp.) using a refractive index increment, dn/dc, of 0.146 mL g -1 [24] .
8
Isolation and Characterization of Grifola frondosa Polysaccharide
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Grifola frondosa polysaccharide (GFI) was isolated from the fruit bodies of G. frondosa. In brief, the dry fruit bodies of G. frondosa were ground to powder. The powder was soaked in deionized water at a ratio of 1:20 (w/v) for 30 minutes, and then extracted by refluxing two times at 100°C. The extract was filtered and concentrated to a small volume. The polysaccharide fraction was precipitated overnight at 4°C by adding 4 volumes of ethanol in the extract. The precipitate was re-dissolved in water, dialyzed against water for 72 hours, concentrated, filtered and dried by a vacuum freeze-drying.
The molar mass distribution of GFI was determined by high-performance gel filtration chromatography (HPGFC) on an Agilent 1200 high-performance liquid chromatography (HPLC) system with differential refraction detector (1260 Infinity, Agilent, USA). After filtered with 0.22 μm filter, the polysaccharide solution (5 mg/mL, 10 μL) was separated in the TSKgel G3000PWXL and TSKgel G5000PWXL columns (7.8 mm × 30 cm, Tosoh Bioscience, Tokyo, Japan), with 0.01 M NaCl elution solution at 0.5 mL/min at 35 °C. The average molecular weight of the polysaccharide ranged from 0.24 to 99.9 x 104Da (Fig. S2 and Table S2 ).
The molar mass distribution of GFI was determined by high-performance gel filtration chromatography (HPGFC) on an Agilent 1200 high-performance liquid chromatography (HPLC) system with differential refraction detector (1260 Infinity, Agilent, USA). After filtered with 0.22 μm filter, the polysaccharide solution (5 mg/mL, 10 μL) was separated in the TSKgel G3000PWXL and TSKgel G5000PWXL columns (7.8 mm × 30 cm, Tosoh Bioscience, Tokyo, Japan), with 0.01 M NaCl elution solution at 0.5 mL/min at 35 °C. The average molecular weight of the polysaccharide ranged from 0.24 to 99.9 x 104Da (
9
Partial Hydrolysis of Chitin-Peptide Complex
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Extracted CPC-P was partially hydrolyzed using 0.1 M or 0.5 M TFA at 70 °C for 3 h to obtain low molecular weight CPC-P. After removing TFA by drying, the hydrolysates were dissolved in distilled water. Partially hydrolyzed CPC-P in 0.1 M and 0.5 M TFA were named SP0.1 and SP0.5, respectively. Molecular weights of CPC-P, DSP, SP0.1 and SP0.5 were determined by size-exclusion chromatography (LC-6A; Shimadzu Co., Kyoto, Japan) equipped with a column of TSKgel G5000 PWXL (7.8 mm × 300 mm, Tosoh Co., Kyoto, Japan) and a refractive index detector RID-10A at 35 °C.28) The column was eluted by water at a flow rate of 1 mL/min. Pullulan P-10 (molecular weight = 0.96 × 104), P-50 (4.71 × 104), P-200 (20.0 × 104) and P-800 (70.8 × 104) (Showa Denko Co., Tokyo, Japan) were used as the molecular weight standards.
10
Gel Permeation Chromatography of SSPS and Oxidized Products
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The molecular weight of SSPS and its oxidized products (DPA) were determined by gel permeation chromatography (GPC) with TSKgelG5000PWXL (Tosoh Corporation, Japan). 0.1 g dried sample was completely dissolved in phosphate buffer saline (PBS) and then filtered using a 0.22 μm pore membrane to eliminate dust particles. The concentration of the samples was 5 mg mL−1, and an injection volume of 20 μL was employed. The mobile phase was 0.1 mol L−1 PBS at a flow rate of 0.6 mL min−1.20 (link)
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