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2 protocols using l leucine d10

1

Metabolite Quantification Using Labeled Standards

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Mass-grade methanol and acetonitrile were from Fisher Chemical (Thermo Fisher Scientific, Boston, MA, USA). Ammonium bicarbonate and formic acid were purchased from Fluka (Dresden, Germany). The ultrapure water was prepared with a Milli Q purification system (Millipore, Burlington, MA, USA). The chemical standards for compound identification were obtained from Sigma-Aldrich (St. Louis, MO, USA), Adamas (Hong Kong, China) or JK Chemical Ltd. (Shanghai, China). The deuterium-labeled internal standards (ISs), including cholic acid-d4,chenodeoxycholic acid-d4, succinic acid-d4,L-leucine-d10, L-phenylalanine-d5, L-tryptophan-d5, L-citrulline-d4, acylcarnitine C10:0-d3 and acylcarnitine C10:0-d3 were from Cambridge Isotope Laboratories, and the natural lipid analogs, including palmitic acid-d3and 1stearic acid-d3,were supplied by Avanti Polar Lipids.
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2

Isotopic Glycoproteomics: Workflow and Methods

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Leucine (L-leucine-13C, L-leucine-13C6,15N, and L-leucine-D10), glycine (glycine-d2 and glycine-13C2,15N), heavy formaldehyde (CD2O), sodium cyanoborodeuteride were purchased from Cambridge Isotope Laboratories, Inc. (Tewksbury, MA). Acetic acid (AA), acetonitrile (ACN), dimethyl sulfoxide (DMSO), formic acid (FA), methanol (MeOH), and water were purchased from Fisher Scientific (Pittsburgh, PA). Sodium cyanoborohydride, sodium meta-periodate, triethylammonium bicarbonate buffer (TEAB, 1.0 M) and tris(2-carboxy-ethyl) phosphine hydrochloride (TCEP) were purchased from Sigma-Aldrich (St. Louis, MO). PNGase F was purchased from Promega (Madison, WI). Oasis HLB 1cm3 cartridges were purchased from Waters Corporation (Milford, MA). Bovine thyroglobulin (BTG) and human serum protein (HSP) were purchased from Thermo Fisher Scientific (Rockford, IL). Microcon-30kDa centrifugal filters (30K MWCO) were purchased from Merck Millipore Ltd. (Darmstadt, Germany). PolyGLYCOPLEX A™ beads (3 μm) were purchased from PolyLC Inc. (Columbia, MD). Fused silica capillary tubing (inner diameter 75 μm, outer diameter 375 μm) was purchased from Polymicro Technologies (Phoenix, AZ). All reagents were used without additional purification.
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