Free xylose levels were quantified using a colorimetric-based D-xylose assay kit (Megazyme). Stool samples were weighed, resuspended in 1 ml ddH2O, vortexed for 30 s, and then centrifuged at 13,000 g for 2 min to remove large particles. Next, supernatants were transferred to new tubes, and the manufacturer’s instructions were closely followed to measure xylose levels, including the step of Carrez purification to remove protein inhibitors.
D xylose assay kit
Manufactured by Megazyme
Sourced in Ireland
The D-xylose assay kit is a laboratory tool designed to quantify the concentration of D-xylose, a naturally occurring sugar, in various samples. It provides a reliable and efficient method for the analysis of D-xylose levels.
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Lab products found in correlation
Enzychrom glucose assay kit, by BioAssay Systems (1 mentions)
Multiskan go uv spectrophotometer, by Thermo Fisher Scientific (1 mentions)
Raffinose d galactose assay kit, by Megazyme (1 mentions)
L rhamnose assay kit, by Megazyme (1 mentions)
Pentra 400, by Horiba (1 mentions)
Dna polymerase, by New England Biolabs (1 mentions)
Dna ligase, by New England Biolabs (1 mentions)
Zymo dna clean and concentrator 5, by Zymo Research (1 mentions)
E z n a yeast dna kit, by Omega Bio-Tek (1 mentions)
Gblock, by Integrated DNA Technologies (1 mentions)
Cellulase from trichoderma reesei atcc 26921, by Merck Group (1 mentions)
Glucose cii test, by Fujifilm (1 mentions)
Silica gel 60 f254, by Merck Group (1 mentions)
Pierce protein assay kit, by Thermo Fisher Scientific (1 mentions)
P nitrophenyl β d xylopyranoside pnpx, by Merck Group (1 mentions)
96 well polystyrene plate, by Corning (1 mentions)
Spectramax i3 spectrometer, by Molecular Devices (1 mentions)
Alliance hplc, by Waters Corporation (1 mentions)
Microq tof mass spectrometer, by Waters Corporation (1 mentions)
Gopod format, by Megazyme (1 mentions)
19 protocols using d xylose assay kit
1
Quantifying Fecal Xylose Levels
2
Glucose, Xylose, and Ethanol Quantification
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The glucose and xylose consumed and the ethanol produced were assayed using the EnzyChrom™ Glucose Assay Kit (BioAssay Systems, Hayward, CA; Catalog #EBGL‐100), the d ‐Xylose Assay Kit (Megazyme, Bray, Ireland), and the Ethanol test kit, respectively, (Thermo Fisher Scientific, Waltham, MA; Catalog #NC9508587) in all cases using methods provided by the manufacturer.
3
Saccharide Profiling of Red Beet Juice
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The saccharide profiles of the red beet juice were analyzed using Megazyme kits (Sucrose Fructose/D-Glucose Assay Kit, Raffinose/D-Galactose Assay Kit, L-Rhamnose Assay Kit, D-Xylose Assay Kit) (Megazyme Inc., County Wicklow, Ireland) and using a Multiskan GO UV spectrophotometer (Thermo Fisher Scientific, Munich, Germany), according to the manufacturer's instructions, as previously described by Modelska et al. [21] . Total sugar content was determined using the Luff-Schoorl method, in accordance with the Polish Standard PN-90/A-75101/07 and the Grain and Feed Trade Association (GAFTA) Method 10. 1. [22,23] . This method is based on hot reduction of an alkaline copper salt solution by direct titration, using a reducing sugar solution in the presence of methylene blue as an indicator. The reduction of the Cu(II) ions present in the Luff solution by the saccharides in the analysed sample was initiated at the boiling point. The volume of sodium thiosulphate (VI) corresponding to the amount of copper (II) reduced by saccharides was calculated as the difference between the volumes obtained from two (blank and specific) titrations. Based on these results, the content of reducing saccharides was determined in each sample [24] (link).
4
D-xylose Absorption Test in Patients
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After a 12 h fast, seven patients ingested 15 g D-xylose dissolved in 150 mL water and after that another 150 mL of water. D-xylose was measured in the blood after 60 min. The plasma concentration of D-xylose was measured with the ‘D-xylose Assay Kit’ from Megazyme© on Pentra 400 (HORIBA ABX SAS).
5
Crude Extract α-Xylosidase Assay
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A preparation of crude extract from seedlings and the α-xylosidase assay were prepared according to Sampedro et al. (2010) (link). XXXG (a gift from Dr Kazuhiko Nishitani) was used as a substrate, and released xylose was quantified using the D-Xylose Assay Kit (Megazyme, Ireland).
6
Molecular Techniques for Yeast Genomics
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All chemicals were purchased from Sigma unless otherwise stated. All restriction enzymes, DNA ligases, and DNA polymerases used for cloning and PCR were purchased from New England Biolabs (NEB) unless otherwise stated. Plasmid minipreps, PCR purifications, and gel extractions were done using the Zymo Zyppy Plasmid Miniprep Kit, and Zymo DNA Clean and Concentrator 5. Genomic DNA from Y. lipolytica was extracted using the E.Z.N.A. Yeast DNA kit (Omega Biotek). All oligonucleotides and gBlocks® were purchased from IDTDNA. Xylose assays were performed using the D-Xylose Assay Kit (Megazyme). Glucose assays were done using glucose oxidase and horseradish peroxidase enzymes purchased from Sigma.
7
Metabolic Evaluation of D-Xylose Absorption
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Urine samples were collected weekly from metabolic cages and centrifuged 10 minutes at 2500 rpm and 4°C (TGL-20MS, Lu Xiangyi Centrifuge Instrument Co., Ltd., Shanghai, China). Then the supernatants were collected. After 4 weeks, rats were given a 3% D-xylose solution by gavage at 10 mL/kg of body weight, except 2 in group C, after 18 hours' fasting. Then the rats were anesthetized with 10% chloral hydrate through intraperitoneal injection at 3 mL/kg of body weight. Blood samples were obtained from the abdominal aorta and centrifuged for 15 minutes at 3000 rpm and 4°C, and then the serum was collected. All samples were stored at −80°C until biochemical analysis.
The serum cortisone, adrenocorticotropic hormone (ACTH), and 24 h urine 17-hydroxycorticosteroid (17-OHCS) levels were measured using ELISA kits (EIAaB Science Co., Ltd., Wuhan, China). The serum D-xylose level was measured with a D-xylose assay kit (Megazyme Ltd., Wicklow, Ireland).
The serum cortisone, adrenocorticotropic hormone (ACTH), and 24 h urine 17-hydroxycorticosteroid (17-OHCS) levels were measured using ELISA kits (EIAaB Science Co., Ltd., Wuhan, China). The serum D-xylose level was measured with a D-xylose assay kit (Megazyme Ltd., Wicklow, Ireland).
8
Quantitative Glucose and Xylose Analysis
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A. nidulans strains were grown in 50 ml minimal media supplemented with 1% (w/v) glucose or 1% (w/v) xylose at 37°C, 150 rpm, for different time periods. At each time point, 5.0 ml of the culture supernatant was harvested. Glucose and xylose concentrations of the supernatants were measured using the Glucose GOD-PAP Liquid Stable Mono-reagent enzymatic kit from LaborLab Laboratories Ltd. (Guarulhos, SP, Brazil) and the D-xylose assay kit from Megazyme. All assays were carried out according to manufacturer’s instructions.
9
Enzymatic Saccharification of Plant Biomass
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The powdered stem samples were treated with a mixture of cellulase from Trichoderma reesei ATCC 26921 (Sigma-Aldrich, Merck KgaA, Darmstadt, Germany) and cellobiase from Aspergillus niger (Sigma-Aldrich, Merck KgaA) for 24 h, according to the methods of Okubo-Kurihara et al. (2016) (link) and Ohtani et al. (2017b) (link). The supernatant was collected after centrifugation, after which 0.1 M NaOH solution was added to stop the reaction. The released glucose and xylose were measured with a “Glucose CII-Test” (FUJIFILM Wako Pure Chemical Corporation, Osak, Japan) and a “D-Xylose Assay Kit” (Megazyme, Bray, Ireland), respectively.
10
Quantifying Reducing Sugars and Ethanol
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Reducing sugars concentration was determined according to dinitro-3.5-salicilic acid (DNS) method [47 (link)]. The effect of ethanol on the sugar measuring method was determined by control measurements, containing all the ethanol concentrations used and a standard sugar concentration. Ethanol appeared to have no effect on the reducing sugar determining method. Glucose was measured according to commercial enzyme solution of GOD/PAP (glucose oxidase/ peroxidase assay) (Biosis, Greece). Xylose was measured according to commercial D-xylose assay kit (Megazyme, Ireland) Ethanol was analyzed using an HPLC system (Szimadju) equipped with an Aminex HPX-87H column (Bio-Rad, 300 x 7.8 mm, particle size 9 μm) using a Refractive Index (RI) detector. Mobile phase was 5 mM H2SO4 in HPLC grade water at 0.6 mL/min flow rate, column temperature was 40°C, injection volume was 50 μl and total runtime was 30 min. All samples were filtered (0,2 μm, Macherey-Nagel) prior to the analysis.
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