The homogeneity and MW determination of AGSP and S-AGSP were conducted using a liquid chromatography instrument, Waters 2695 (Waters Co., Milford, MA, USA), equipped with a 7.8 × 300 mm TSK-gel G4000PWXL column (Tosoh Bioscience, Tokyo, Japan), and a 2414 refractive index detector (Waters Co., Milford, MA, USA), respectively. A sample solution of 20 µL (2 mg/mL) was injected into the system for each run. Ammonium acetate solution (0.1 mol/L) was used as eluent at a flow rate of 0.4 mL/min. The column was previously calibrated using standard dextrans, including 1000, 5000, 25,000, 50,000, 150,000, 410,000 and 670,000 Da.
Tsk gel g4000pwxl column
Manufactured by Tosoh
Sourced in Japan, United States
The TSK gel G4000PWXL column is a gel permeation chromatography (GPC) column designed for the analysis of high molecular weight polymers. The column is packed with a porous polymer-based material that separates molecules based on their size and molecular weight. The column can be used to determine the molecular weight distribution of various types of polymers.
Automatically generated - may contain errors
Lab products found in correlation
Lc 10atvp hplc system, by Shimadzu (2 mentions)
2414 refractive index detector, by Waters Corporation (1 mentions)
Waters 2695, by Waters Corporation (1 mentions)
Milli q synthesis, by Merck Group (1 mentions)
Agilent 1260, by Agilent Technologies (1 mentions)
Dawn heleos 2, by Wyatt Technology (1 mentions)
Optilab rex, by Wyatt Technology (1 mentions)
Thermo ultimate 3000 system, by Thermo Fisher Scientific (1 mentions)
Benzonase, by Merck Group (1 mentions)
Chloroform, by Merck Group (1 mentions)
Lc 10avp system, by Shimadzu (1 mentions)
Malvern zetasizer nano zs90, by Malvern Panalytical (1 mentions)
Dextrans, by Merck Group (1 mentions)
Optilab t rex, by Wyatt Technology (1 mentions)
Agilent 1260 infinity, by Agilent Technologies (1 mentions)
Zorbax eclipse xdb c18 column, by Agilent Technologies (1 mentions)
Waters 2998, by Waters Corporation (1 mentions)
Lc 10advp, by Shimadzu (1 mentions)
1260 rid detector, by Agilent Technologies (1 mentions)
Pepsin, by Merck Group (1 mentions)
18 protocols using tsk gel g4000pwxl column
1
Determination of AGSP and S-AGSP Homogeneity and MW
2
Molecular Weight Analysis of CP Sample
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The molecular weight of the CP sample was analyzed using an HPLC apparatus (Agilent 1260, Agilent Technologies, Walnut Creek, CA, USA) equipped with DAWN HELEOS-II (Wyatt Technology Corporation, America) and Optilabr EX (Wyatt Technology Corporation, USA) detectors coupled with a TSK gel G4000PWxl column (7.8 × 300 mm, TOSOH, Tokyo, Japan), following the previous study (25 (link)). The mobile phase included 0.1% trifluoroacetic acid, 45% acetonitrile, and 54.9% ultrapure water (ultrapure water machine, Milli-Q Synthesis, Millipore Corporation, USA). The standard was composed of Gly-Sar (146 Da), Gly-Gly-Tyr-Arg (451 Da), bacitracin (1,422 Da), aprotinin (6,511 Da), and cytochrome C (12,327 Da).
3
Polysaccharide Molecular Weight Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The homogeneity and molecular weights of RS-3-1, RS-3-2, BZ-3-1, BZ-3-2, BZ-3-3, FL-3-1, and GC-3-1 were evaluated and determined by high performance gel permeation chromatography (HPGPC) on a TSK-GEL G4000PWxl column (7.5 × 300 mm, Tosoh Co., Japan). The columns were maintained at 30°C, and the mobile phase was deionized water at a flow of 1 mL/min. T-series dextrans with different molecular weights (1 × 104, 2 × 104, 4 × 104, 7 × 104, 1.1 × 105 Da) were used for the calibration curve. A 20 μL aliquot was injected for each run. A standard linear curve was calibrated with dextran standards, Y = −0.0198X + 4.8426 (R2 = 0.9900). The molecular weight of these polysaccharides were estimated using the calibration curve equation.
4
PNPB1 Molecular Characterization by HPGPC
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The homogeneity and molecular weight of PNPB1 were determined by high-performance gel permeation chromatography (HPGPC) using a Thermo Ultimate 3,000 system (Thermo Fisher Scientific Co., United States) equipped with an Alltech 2000 ES evaporative light scattering detector (ELSD) and a TSK gel G4000PWXL column (7.8 × 300 mm, Tosoh Corp, Tokyo, Japan). The sample solution (1.0 mg/mL, 10 μL) was injected and eluted with distilled water at 30°C with a flow rate of 1.0 mL/min (Qiao et al., 2010 (link)). The molecular weight of PNPB1 was estimated by reference to the calibration curve established by Dextran standards of known molecular weight (80 kDa, 150 kDa, 270 kDa, 410 kDa, 670 kDa and 990 kDa).
5
Quantification of FMD Virus Particles
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The amount of the FMD vaccine antigen (146S) was measured as previously described [13 (link)]. Briefly, the viral infection supernatant was treated with chloroform (Merck) at a ratio of 1:1 (v/v) and mixed vigorously for 5 min. The mixture was then centrifuged at 4000× g for 15 min at 4 °C, and then the aqueous phase on top of the organic solvent was collected. Next, the samples were treated with benzonase (Sigma-Aldrich, St. Louis, MO, USA) at a concentration of 0.025 units/μL, and they were incubated at 37 °C for 1 h with shaking. Then, the samples were clarified by centrifugation at 16,000× g for 10 min at 4 °C. The FMDV intact particles in the samples were measured by loading the samples onto a high-performance liquid chromatography (Agilent Technologies, Santa Clara, CA, USA) fitted with a TSKgel G4000PWXL column (TOSOH Bioscience, Tokyo, Japan). Finally, the peak area was integrated for the quantification of FMDV particles.
6
Pectin Extraction and Characterization from Apple Peel
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Pectin content in apple peel was determined by a carbazole sulfuric acid method as described previously (Liang et al., 2022 (link)). For its structural characterization, the pectin material was extracted from apple peel according to the method of Pasandide et al. (2017) (link) with some modifications. Briefly, apple peel powder (4.0 g) was extracted twice with 0.05 M hydrochloric acid solution (60 mL) at 80 °C for 2 h with continuous stirring. After each extraction, the slurry was centrifuged at 4000 rpm for 20 min. The obtained supernatants were combined and the molecular weight distribution of polysaccharides was determined by a high-performance size exclusion chromatography (HPSEC) equipped with a TSK gel G4000PWXL column (TOSOH, Tokyo, Japan) and a refractive index RID-10A detector (Shimadzu, Tokyo, Japan) (Feng et al., 2019 (link)). Moreover, the supernatant (50 mL) was mixed with absolute ethanol (1:1, v/v) for 6 h at ambient temperature and the precipitate was collected by a centrifugation at 4000 rpm for 15 min representing the higher molecular weight fraction of pectin material. Upon lyophilization, its monosaccharide composition was analyzed as described by Feng et al. (2019) (link).
7
Molecular Weight Determination of WGP
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
WGP (5 mg/ml) was filtered via 0.45 μm microfiltration membrane. The filtered sample (20 μL) was loaded into a TSK-Gel G4000PWXL column (Tosoh, Shanghai branch, China) controlled by LC-10Avp system (Shimadzu, Shanghai branch, China). High performance gel permeation chromatography (HPGPC) was performed using 0.2 M NaCl as mobile phase at flow rate of 0.5 ml/min. T-series Dextran standards were used for reference standards.
8
Molecular Weight and Particle Size Analysis
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The molecular weight was analyzed by LC-10ATvp HPLC system (Shimadzu, Tokyo, Japan), which is equipped with a TSK-GEL G4000PWXL column (Tosoh Co., Tokyo, Japan) and an Alltech 2000ESELSD (Shimadzu, Tokyo, Japan). D.D. water was served as the mobile phase, the flow rate was 0.45 mL/min, aerosol level was 60%, drift tube temperature was 120 °C and nebulizing nitrogen pressure was 25 psi. The dextran standards were used to create a calibration curve [34 (link)]. The particle size of polysaccharides in deionized water was detected by Dynamic light scattering (DLS) using a Malvern Zetasizer Nano (ZS90, Malvern Instruments Ltd., Malvern, UK). The sample was dissolved in the D.D. water at a concentration of 1 mg/mL, and there were 10 runs performed in each procedure.
9
Molecular Weight Determination of PTA
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An LC-10ATvp HPLC system (Shimadzu, Kyoto, Japan) equipped with an Alltech 2000 ES evaporative photodetector (Nicholasville, Kentucky, USA) and a TSK-GEL G4000PWXL column (7.8 × 300 mm, Tosoh, Tokyo, Japan) was used to analyze the molecular weight of PTA. It was operated at 35 °C and eluted by ultrapure water at a flow rate of 0.45 mL/min with an initial injection of 20 μL of PTA sample dissolved in ultrapure water. A standard curve established using dextrans of 1, 5, 12, 25, 50 and 80 kDa (Sigma-Aldrich, St. Louis, Missouri, USA) was used to calculate the molecular weight of PTA.
10
SEC-MALS-UV-RI Analysis of Anti-ST1 mAb-Polysaccharide Complexes
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
The SEC-MALS-UV-RI analysis was performed in the method and chromatography system below. The chromatography condition was the same to the Chromatography condition-A in Supplementary Table 12 , except a Tosoh TSKgel G4000PWxL column (7.8 × 300 mm) was used. A multi-angle light scattering (MALS) detector (MiniDAWN®) and a refractive index (RI) detector (Optilab® T-rEX) (Wyatt Technology Corp., Santa Barbara) were added to the system and connected in tandem with the UV detector. All data were collected and analyzed with ASTRA 7 software. A first-order fit Zimm formalism with Astra protein conjugate method was used for Mw calculations47 (link)–49 (link).
Anti-ST1 mAb was complexed with ST1 polysaccharide (Ps) standard at different mAb/Ps ratio as in Supplementary Table13 . The binding reactions were incubated at ambient temperature for 2 h and injected on the SEC-MALS-UV-RI system for analysis.
Anti-ST1 mAb was complexed with ST1 polysaccharide (Ps) standard at different mAb/Ps ratio as in Supplementary Table
About PubCompare
Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.
We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.
However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.
Ready to get started?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!