Trimethylsiloxy-terminated PDMS (silicone oil) with a molecular weight of 5970 g/mol was purchased from Gelest (Morrisville PA, USA). Glass spheres (GS), montmorillonite (MMT), and kaolin (K) were purchased from Sigma Aldrich. Aeroperl 300 Pharma colloidal silica (aero) was kindly supplied by Evonik (Istanbul, Turkey) via Marmara Ecza (Istanbul, Turkey). HDK N20 fumed silica was kindly supplied by Wacker (Istanbul, Turkey) via IMCD Group (Istanbul, Turkey). Whatman Grade 1 filter paper (WFP) was purchased from General Electric (Turkey). Reagent grade tetrahydrofuran (THF) and isoproponal (IPA) were purchased from Merck (Germany) and used as received.
Glass spheres
Manufactured by Merck Group
Glass spheres are small, round particles made of glass. They serve as a basic component in various laboratory equipment and apparatus.
Automatically generated - may contain errors
Lab products found in correlation
Kaolin, by Merck Group (1 mentions)
Aeroperl 300 pharma colloidal silica, by Evonik (1 mentions)
Reagent grade tetrahydrofuran thf, by Merck Group (1 mentions)
Montmorillonite mmt, by Merck Group (1 mentions)
4 nitrophenyl β d glucuronide p npg, by Thermo Fisher Scientific (1 mentions)
β glucosidase from aspergillus niger, by Merck Group (1 mentions)
Na2pdcl4, by Merck Group (1 mentions)
Silica gel, by Merck Group (1 mentions)
Thermomixer, by Eppendorf (1 mentions)
Ethanol, by Merck Group (1 mentions)
6 protocols using glass spheres
1
Silicone oil-based composite preparation
2
Immobilized Enzyme System for Mogroside V
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Lo Han Kuo extract powder (Mogroside V ≥25.0%) was purchased from Huitong bio‐technology company (Guangxi, China). β‐Glucosidase from Aspergillus niger was purchased from Sigma‐Aldrich (St. Louis, MO). 4‐Nitrophenyl β‐D‐glucuronide (pNPG) purchased from Alfa Aesar (Ward Hill, MA) was used as the substrate to determine the catalytic activity of immobilized enzyme system. Glass spheres, nylon pellets, and cellulose beads were separately utilized as solid carriers for enzyme immobilization. Glass spheres were purchased from Sigma‐Aldrich with 9–13 μm particle size. Nylon pellets were provided by Keen Corp Company (Tainan, Taiwan). Porous cellulose beads were synthesized as previously described (Chen & Tsao, 1976 ). The particle size of nylon pellets and cellulose beads was about 1 mm.
3
Phagocytosis by NKTert Cells
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We tested whether the presence of eat-me signals in general is a requirement for phagocytosis by NKTert. For this, we cultured CellTracker Blue labeled NKTert cells for 24 hours in wells of a 384 Greiner μClear plate, before addition of uncoated glass beads (Sigma Aldrich glass spheres 9- to 13-μm particle size; KatNr.: 440345-100g) washed twice with PBS. After 16 h, the cultures were washed twice with RPMI + 10 % FBS to remove non-phagocytosed beads. After staining with lysosomal dye NIR, the cells were imaged on an Opera Phenix microscope in confocal mode.
4
Synthesis of Heterogeneous Catalyst
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Lithium pellets (99%), naphthalene (99%), 1.0 M di-n-butylmagnesium (MgBu2) in heptane, anhydrous tetrahydrofuran (THF), anhydrous isopropanol (IPA), polyvinylpyrrolidone (PVP) (average mol. weight 10 000), Na2PdCl4 (99.99%), silica gel (high-purity, 60 Å, 60–100 mesh), and glass spheres (9–13 μm) were purchased from Sigma-Aldrich. Poly(styrene-b-4-vinyl pyridine) (PS-P4VP) was purchased from Polymer Source, Inc. A gas mixture containing 5 vol% of acetylene in nitrogen was purchased from BOC.
5
Visualization of Prion Protein Binding
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To assess whether recombinant PrP and/or misfolded rec-PrP could be bound to the glass beads used in PMSA reactions, glass spheres from 9 to 13 µm of diameter (Sigma-Aldrich) were chosen. Five different samples were prepared for visualization through SEM: glass spheres washed with distilled water, glass spheres incubated for 1 h at 37 °C and 700 rpm (Thermomixer, Eppendorf) with substrate devoid of rec-PrP, glass spheres incubated in the same conditions with bank vole rec-PrP containing PMSA substrate, glass spheres submitted to PMSA for four 24 h serial rounds, seeded 1:10 in the first one with Ust02 recombinant prions, letting spheres sediment for 15 min after each round and replacing the supernatant for fresh PMSA substrate. After the last PMSA round, glass spheres were harvested and divided in to parts, submitting one half to proteinase K digestion at 5 µg/ml for 1 h at 42 °C and 450 rpm. Both, PK-treated and untreated glass spheres were thoroughly washed with distilled water, and as all the rest of spheres were oven-dried before preparing for visualization. Once all dried, they were sputtered with gold at 25 mA (Emitech K550X, Aname) and analysed by secondary electron emission with a Hitachi S-4000 SEM, operated at 5 kV (SGIker, UPV/EHU).
6
Prednisone Dissolution Analysis
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