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3 protocols using d galactose

1

Glycan Synthesis and Characterization Protocol

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l-fucose was purchased from AppliChem (Darmstadt, Germany). Methyl α-l-fucopyranoside, d-glucose, and d-galactose were purchased from Carbosynth (Compton, UK). 3-O-methyl-d-glucose, biotin, and streptavidin were purchased from Sigma-Aldrich (St. Louis, MO, USA). biotinylated α-l-fucoside was purchased from Synthaur LLC (Moscow, Russia). Methylated disaccharide 3,6-O-Me2-d-Glcβ1-4(2,3-O-Me2)-l-Rhaα-O-(p-C6H4)-O-CH2CH2NH2 was synthesized as described previously [29 (link)]. DyLight 488 NHS ester was purchased from ThermoScientific (Rockford, MI, USA). Protein molecular Marker III was purchased from AppliChem (Darmstadt, Germany). Other chemicals were purchased from Sigma-Aldrich, Duchefa, ForMedium and Applichem companies.
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2

Saccharide Synthesis and Characterization

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Methyl-α-l-fucopyranoside, methyl-β-l-fucopyranoside, d-glucose and d-galactose were purchased from Carbosynth, Compton, United Kingdom; blood group A/B/O trisaccharides, and Fucα1-GlcNAc and Fucα1-GlcNAc were purchased from Carbohydrate Synthesis, Oxford, United Kingdom; l-fucose was purchased from Applichem, Darmstadt, Germany. N-acetyl-d-glucosamine, d-mannose, d-mannose-agarose, biotin and streptavidin, bovine serum albumin, 3,4-dihydroxy-dl-phenylalanine, zymosan A from Saccharomyces cerevisiae and luminol were purchased from Sigma-Aldrich, St Louis, USA. Biotinylated saccharides (biotinylated α/β-d-mannoside, α/β-d-galactoside and α-l-fucoside) were purchased from Synthaur LLC, Moscow, Russia. Fluorescein isothiocyanate (FITC) and DyLight 488 were purchased from ThermoScientific, Rockford, USA. Basic chemicals were purchased from Sigma-Aldrich, St Louis, USA; Duchefa, Haarlem, Netherlands; ForMedium, Hunstanton, United Kingdom and Applichem, Darmstadt, Germany.
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3

Glycosylation Reactions under Inert Atmosphere

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All reactions were performed under an inert atmosphere of nitrogen or argon, unless otherwise noted. d-Galactose, d-glucosamine hydrochloride, and l-fucose were purchased from Carbosynth LLC (CA, USA). All other reagents were purchased from commercial sources and used directly. All solvents were dried and distilled following standard protocols. All glycosylation reactions were performed in an oven-dried round-bottom flask. Proton nuclear magnetic resonance (1H NMR) and 13C NMR spectra were recorded with a Varian 400 MHz spectrometer and a Bruker 600 MHz spectrometer. High resolution mass spectra (HRMS) were acquired using an UltraFlex II MALDI/TOF mass spectrometer (Bruker Corporation, MA, USA) and Orbitrap Lumos mass spectrometer (Thermo Fisher Scientific, CA, USA). Thin layer chromatography (TLC) was performed on a silica gel matrix with a 254 nm fluorescent indicator, and flash column chromatography purification was performed on silica gel 60 (Sigma-Aldrich Corporation, WI, USA).
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