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2 protocols using fmoc rink amide protide non preloaded resin

1

Synthetic Bioconjugation Protocol

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All materials were used according to the manufacturer’s instructions unless otherwise noted. The following were purchased from Sigma Aldrich: acetonitrile (MeCN), methanol (MeOH), triethylamine (TEA), dimethylsulfoxide (DMSO), diethyl ether (ET2O), cyanocobalamin (B12), triisopropylsilane (TIPS), 1,1’-carbonyl-di-(1,2,4-trizole) (CDT), n-methyl-pyrrolidone (NMP), 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDC), 1-hydroxybenzo-triazole (HOBt), and dihydroxybenzoic acid (DHB). The following were purchased from CEM Corporation: Fmoc-Rink Amide Protide non-preloaded Resin (LL), N,N′-diisopropylcarbodiimide (DIC), ethyl cyanohydroxyiminoacetate (Oxyma), Fmoc protected amino acids: Asn(Trt); Gln(OtBu); Tyr(tBu); and Trp(Boc). The following were purchased from VWR:4-trifloroacetic acid (TFA), dimethylformamide (DMF). The following was purchased from BroadPharm: sulfo-DBCO-amine. The following was purchased from ThermoFisher: AlexaFluor®564 (DBCO-AF546). The following was purchased from Lumiprobe: Sulfo-cyanine5 NHS ester. The following was purchased from Acros Organics: alpha-Cyano-4-hydroxycinnamic acid (CHCA).
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2

Peptide Synthesis and Labeling Protocol

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All materials were used according to the manufacturer's instructions unless otherwise noted. The following were purchased from Sigma‐Aldrich, St. Louis, MO, USA: acetonitrile (MeCN), methanol (MeOH), triethylamine (TEA), dimethyl sulphoxide (DMSO), diethyl ether, cyanocobalamin (B12), triisopropylsilane, 1,1′‐carbonyl‐di‐(1,2,4‐trizole), n‐methyl‐pyrrolidone, 1‐ethyl‐3‐(3‐dimethylaminopropyl) carbodiimide, 1‐hydroxybenzo‐triazole and dihydroxybenzoic acid. The following were purchased from CEM Corporation, Matthews, NC, USA: Fmoc‐Rink Amide Protide non‐preloaded resin (LL), N,N′‐diisopropylcarbodiimide, ethyl cyanohydroxyiminoacetate (Oxyma), Fmoc protected amino acids: Asn(Trt); Gln(OtBu); Tyr(tBu); and Trp(Boc). The following were purchased from VWR, Radnor, PA, USA: 7‐trifloroacetic acid (TFA) and dimethylformamide (DMF). The following was purchased from BroadPharm, San Diego, CA, USA: sulpho‐DBCO‐amine. The following was purchased from Thermo Fisher, Waltham, MA, USA: AlexaFluor 564 (DBCO‐AF546). The following was purchased from Lumiprobe: Sulfo‐cyanine5 NHS ester. The following was purchased from Lumiprobe, Hunt Valley, MD, USA: alpha‐cyano‐4‐hydroxycinnamic acid.
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