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2 protocols using cysta

1

Comprehensive Metabolite Quantification Protocol

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Met, SAM, SAH, MTA, Hcy, Cysta, Cys, GSH, 5-MTHF, VB6, VB9, VB12, LC-MS-grade acetonitrile (ACN), LC-grade methanol (MeOH), formic acid, ascorbic acid, sodium hydroxide (NaOH) 0.1 M, hydrochloric acid (HCl), ammonium acetate (NH4OAc), tris-(2-carboxyethyl)phosphine (TCEP), L-Met, and DL-Met were purchased from Sigma-Aldrich (St. Louis, MO, USA). DL-HMTBA was purchased from Macklin (Shanghai, China). DL-MM was purchased from Taopu (Shanghai, China). Homocystine-d8 was purchased from CDN Isotopes (Pointe-Claire, Quebec, Canada). Ultrapure water was obtained from a Millipore-Q water system (Millipore, Bedford, MA, USA). Dulbecco's modified Eagle's medium and Ham's F-12 nutrient mixture (DMEM/F12), RPMI-1640 medium, fetal bovine serum (FBS), and penicillin–streptomycin were purchased from Gibco (Shanghai, China).
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2

Quantification of One-Carbon Metabolites

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Hcy, Met, Cysta, SAH and SAM were purchased from Sigma Aldrich (St. Louis, Missouri, USA). 5m-THF was purchased from Caymen Chemical (Ann Arbor, Michigan, USA). SAM-d3, Hcy-d4, Met-d4 and SAH-d4 were purchased from Toronto Research Chemicals (Toronto, Canada). Cysta-d4 and 5m-THF-d3 were purchased from 13C Molecular, Inc (Fayetteville, North Carolina, USA). Optima liquid chromatography–mass spectrometry grade water (H2O), acetonitrile and formic acid were purchased from Fisher Scientific (Pittsburg, Pennsylvania, USA).
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