ACS-grade solvents (acetone, methanol, and 95% ethanol), HPLC-grade solvents (water, methanol, and acetonitrile), glass wool, Gibco fetal bovine serum, glacial acetic acid and hydrochloric acid were acquired from the Fisher Scientific Co., LLC (Suwanee, GA, USA). Sephadex LH-20, Amberlite XAD-16, Folin & Ciocalteu’s phenol reagent, gallic acid (>99%), (+)-catechin hydrate (>99%), chlorogenic acid (≥98%), protocatechuic acid (≥97%), vanillic acid (≥97%), fluorescein 3′,6′-dihydroxyspiro[isobenzofuran-1[3H]9′[9[H]xanthen]-3-one), Trolox [(±)-6-hydroxy-2,5,7,8-tetramethylchroman-2-carboxylic acid, 97%], TPTZ (2,4,6-tripyridyl-S-triazine, ≥99.0%), quercetin-3-O-rutinoside hydrate (≥94%), iron(II) sulfate heptahydrate (≥99.0%), iron(III) chloride hexahydrate (97%), 2′,7′-dichlorofluorescin diacetate (DCFH-DA, ≥97%), and 2,2′-azobis [2-amidinopropane] dihydrochloride (ABAP) were purchased from the Sigma-Aldrich Chemical Company (St. Louis, MO, USA). Cyanidin-3-O-glucoside chloride was obtained from the Indofine Chemical Company, Inc. (Hillsborough, NJ, USA). Advanced DMEM, phosphate-buffered saline (PBS), Hanks’ balanced salt solution (HBSS), penicillin-streptomycin, and trypsin-EDTA were purchased from Life Technologies (Grand Island, NY, USA). Human colorectal adenocarcinoma (Caco-2) cells were procured from the American Type Culture Collection (ATCC, Manassas, VA, USA).
Glass wool
Manufactured by Thermo Fisher Scientific
Sourced in Belgium, United States
Glass wool is a fibrous material made from extremely fine glass fibers. It is used as a thermal and acoustic insulation material in various applications.
Automatically generated - may contain errors
Lab products found in correlation
Sorvall lynx 6000 superspeed centrifuge, by Thermo Fisher Scientific (2 mentions)
S phenol reagent, by Merck Group (1 mentions)
Iron 2 sulfate heptahydrate, by Merck Group (1 mentions)
2 2 azobis 2 amidinopropane dihydrochloride abap, by Merck Group (1 mentions)
Acs grade solvents, by Thermo Fisher Scientific (1 mentions)
Amberlite xad 16, by Merck Group (1 mentions)
Quercetin 3 o rutinoside hydrate, by Merck Group (1 mentions)
Gibco fetal bovine serum, by Thermo Fisher Scientific (1 mentions)
Hank s balanced salt solution hbss, by Thermo Fisher Scientific (1 mentions)
Sephadex lh 20, by Merck Group (1 mentions)
Penicillin streptomycin, by Thermo Fisher Scientific (1 mentions)
Trypsin edta, by Thermo Fisher Scientific (1 mentions)
Advanced dmem, by Thermo Fisher Scientific (1 mentions)
Iron 3 chloride hexahydrate, by Merck Group (1 mentions)
Hplc grade solvents, by Thermo Fisher Scientific (1 mentions)
2 7 dichlorofluorescin diacetate dcfh da, by Merck Group (1 mentions)
Chlorogenic acid, by Merck Group (1 mentions)
Protocatechuic acid, by Merck Group (1 mentions)
Vanillic acid, by Merck Group (1 mentions)
Phosphate buffered saline (pbs), by Thermo Fisher Scientific (1 mentions)
5 protocols using glass wool
1
Colorectal Cancer Cell Antioxidant Assays
2
Biofilm Cell Isolation and Extraction
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Biofilm cells were collected as previously described with minor modifications56 (link). Overnight cultures of the BW25113 and isogenic prophage deletion strains were inoculated into a 2 L shake flask (to reach OD600 ~ 0.05) containing 250 mL LB and 10 g glass wool (Corning Glass Works, Corning, NY), during which time the cells formed a biofilm on the surface of the glass wool. After incubation with shaking at 180 rpm at 37 °C for 48 h, the planktonic cells in contact with biofilms were taken directly from the culture. At the same time, the glass wool was taken from the culture and quickly washed twice in 100 mL ice-cold 0.85% NaCl. The biofilm cells were removed from the glass wool by sonication at 22 Watts for 2 min in a FS3 water bath sonicator (Fisher Scientific, Pittsburgh, PA, USA) submerged in 200 mL of ice-cold 0.85% NaCl. The biofilm cells were collected by centrifugation at 4 °C for 30 s to remove the NaCl. The cell pellets from the planktonic state growth and biofilm growth were saved at -80 °C until used.
3
Tracing Fungal Nutrient Transport
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At the time of planting, three windowed PVC cores lined with 35 μm nylon mesh (PlastOk Ltd., Birkenhead, UK) were inserted into the pot substrate (Figure S1 A). Mesh, affixed to sides and base of the cores using Tensol® 12 acrylic adhesive (Bostik Ltd, Staffordshire, UK), excluded roots of cv. Skyfall plants but permitted access of extra-radical fungal hyphae [65 ]. Two of the cores were filled with bulk substrate (99.25% core volume) and fine-ground tertiary basalt (0.75% core volume) that acted as fungal bait [22 (link)]. A silicone capillary tube (Smith Medical Inc., Kent, UK) was attached centrally to these cores, via which 33P was later introduced in an aqueous solution to one core in each pot (see 33P isotope tracing). The third core was filled with glass wool (Acros Organics, Geel, Belgium) and fitted with a Suba-Seal® rubber septum (Sigma-Aldrich, Darmstadt, Germany). This core allowed for the sampling of below-ground respiration and flux of 14C by the extra-radical mycelium of the AM fungus throughout the 14C labeling period (see 14C label).
4
Cheese Calcium and Nitrogen Extraction
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A water-soluble extract (WSE) of the cheese was prepared by blending distilled water (50°C) and grated cheese at a weight ratio of 2:1 (Stomacher, Lab-Blender 400; Seward Medical, London, UK) for 5 min, holding at 50°C for 1 h, centrifuging at 3000 g for 20 min at 4°C (Sorvall LYNX 6000 Superspeed centrifuge, Thermo Scientific, Dublin, Ireland), and filtering through glass wool (Acros organics, Geel, Belgium). A portion (4 mL) of filtrate (WSE) was ashed at 550°C and the ash was analyzed for calcium by flame atomic absorption spectroscopy (ISO 8070, IDF 119:2007). Serumsoluble calcium was expressed as a percentage of the total cheese calcium content. A further portion (60 ml) of the WSE was adjusted to pH 4.6 using 10% w/w HCl (Honeywell Fluka™ Chemicals, Offenbach, Germany), centrifuged at 3000 g for 20 minutes at 4°C and filtered through glass wool.
The resultant pH 4.6 soluble filtrate was analyzed for N using the macro-Kjeldahl method (ISO 8968-1:2014, IDF 20-1:2014) and expressed as a percentage of total cheese nitrogen.
Measurements were performed in duplicate per cheese.
The resultant pH 4.6 soluble filtrate was analyzed for N using the macro-Kjeldahl method (ISO 8968-1:2014, IDF 20-1:2014) and expressed as a percentage of total cheese nitrogen.
Measurements were performed in duplicate per cheese.
5
Cheese Calcium and Nitrogen Extraction
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A water-soluble extract (WSE) of the cheese was prepared by blending distilled water (50°C) and grated cheese at a weight ratio of 2:1 (Stomacher, Lab-Blender 400; Seward Medical, London, UK) for 5 min, holding at 50°C for 1 h, centrifuging at 3000 g for 20 min at 4°C (Sorvall LYNX 6000 Superspeed centrifuge, Thermo Scientific, Dublin, Ireland), and filtering through glass wool (Acros organics, Geel, Belgium). A portion (4 mL) of filtrate (WSE) was ashed at 550°C and the ash was analyzed for calcium by flame atomic absorption spectroscopy (ISO 8070, IDF 119:2007). Serumsoluble calcium was expressed as a percentage of the total cheese calcium content. A further portion (60 ml) of the WSE was adjusted to pH 4.6 using 10% w/w HCl (Honeywell Fluka™ Chemicals, Offenbach, Germany), centrifuged at 3000 g for 20 minutes at 4°C and filtered through glass wool.
The resultant pH 4.6 soluble filtrate was analyzed for N using the macro-Kjeldahl method (ISO 8968-1:2014, IDF 20-1:2014) and expressed as a percentage of total cheese nitrogen.
Measurements were performed in duplicate per cheese.
The resultant pH 4.6 soluble filtrate was analyzed for N using the macro-Kjeldahl method (ISO 8968-1:2014, IDF 20-1:2014) and expressed as a percentage of total cheese nitrogen.
Measurements were performed in duplicate per cheese.
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