The A2G2 (NA2) N-glycan, 13C6-aniline, and sodium cyanoborohydride (NaBH3CN), were purchased from (Sigma Aldrich, Milwaukee, WI, USA). The endoglycosidase Endo S was purchased from New England Biolabs (Ipswich, MA, USA). Hypersep Hypercarb PGC SPE cartridges (25 mg, 1 mL) and Pierce BCA Protein Assay Kit were purchased from ThermoFisher Scientific, Waltham, MA, USA). The Sep-PAK C18 solid phase extraction (SPE) cartridges (100 mg, 1 mL) was from Waters (Milford, MA, USA). All other chemicals were of reagent grade or LC/MS grade for chromatography systems. Human recombinant β-Galactosidase was prepared as described [25 (link)].
13c6 aniline
Manufactured by Merck Group
Sourced in United States
13C6-aniline is a chemical compound used as a labeled precursor in various research and analytical applications. It contains six carbon-13 isotopes, which can be used to track the fate and metabolism of aniline in biological systems. The product is typically used in analytical techniques, such as mass spectrometry, to study chemical and biochemical processes.
Automatically generated - may contain errors
Lab products found in correlation
Hypersep hypercarb pgc spe cartridges, by Thermo Fisher Scientific (2 mentions)
Pierce bca protein assay kit, by Thermo Fisher Scientific (2 mentions)
Nabh3cn, by Merck Group (1 mentions)
Seppak c18 solid phase extraction cartridges, by Waters Corporation (1 mentions)
Endo s, by New England Biolabs (1 mentions)
Creatinine assay kit, by Merck Group (1 mentions)
Sodium cyanoborohydride nabh3cn, by Merck Group (1 mentions)
3 protocols using 13c6 aniline
1
Glycosidic Bond Cleavage and Labeling
2
N-glycan Labeling and Purification
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The A2G0 N-glycan was purchased from Prozyme (Hayward, CA). Endoglycosidase Endo S was purchased from New England Biolabs (Ipswich, MA, USA). Natural abundance aniline and [13C6] aniline, sodium cyanoborohydride (NaBH3CN), were purchased from Sigma Aldrich (Milwaukee, WI, USA). Hypersep Hypercarb PGC SPE cartridges (25 mg, 1 mL) and Pierce BCA Protein Assay Kit were purchased from ThermoFisher Scientific (Waltham, MA, USA). Creatinine Assay Kit was purchased from Sigma (St. Louis, MO, USA). The ganglioside standards (GM1, GA1, GM2, GA2 and GM3) were all purchased from Enzo Life Sciences (Farmingdale, NY, USA) while the BMP(14:0) phospholipid was purchased from Avanti Polar Lipids (Alabaster, AL, USA). The 10kDa centrifugal filters were from VWR (Radnor, PA, USA). All other chemicals were of reagent or LC-MS grade for chromatography systems.
3
N-Glycan Labeling and Purification
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Ten μL [12C6]- or [13C6]-aniline (Sigma-Aldrich) and 25 μL fresh NaCNBH3 (1 M) prepared in DMSO/ acetic acid (7:3, v/v) were added separately to amidated N-linked glycans from KK47 and HCV29 cells, and incubated at 75 °C for 10 min as described previously [32 (link), 35 (link)]. The mixture was lyophilized under vacuum, redissolved in 500 μL 1-butanol/ methanol/ H2O (BMW), and desalted with Sepharose 4B. Eluted glycan derivatives were dried and stored in the dark at − 20 °C.
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