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4 protocols using triisopropylsilane tis

1

Peptide Synthesis and Fluorescent Labeling

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Rink
amide resin (200–400 mesh, loading = 0.345 mmol/g), Fmoc-Gly-OH,
Fmoc-Pro-OH, Fmoc-Hyp(tBu)-OH, Fmoc-Asp(Otbu)-OH, Fmoc-Tyr(tBu)-OH,
Fmoc-Ala-OH, Fmoc-Phe-OH, O-Benzotriazole-N,N,N′,N′-tetramethyl-uronium-hexafluorophosphate (HBTU),
and N-hydroxybenzotriazole (HOBt, anhydrous) were
purchased from GL Biochemical Company (Shanghai, China). N,N-Diisopropylethylamine (DIEA) was obtained from
Hanhong Chemical Technology Co. Ltd (Shanghai, China). 5(6)-Carboxylfluorescein
(FAM) and piperazine (anhydrous) were provided by Aladdin Industrial
Corporation (Shanghai, China). Trifluoroacetic acid (TFA) and triisopropylsilane
(Tis) were purchased from Tokyo Chemical Industry Co. Ltd (Tokyo,
Japan). Bovine serum albumin (BSA), goat serum, and 4′,6-diamidino-2-phenylindole
(DAPI) were obtained from Solarbio Science & Technology Co. Ltd
(Beijing, China). Hemoglobin, trypsin, and pepsin were obtained from
Yuanye Biological Technology Co. Ltd (Shanghai, China). Gelatin was
obtained from Biotop Science & Technology Co. Ltd (Beijing, China).
All of the commercial reagents were of analytical grade and were used
without further purification.
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2

Peptide Synthesis Reagents and Materials

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Fmoc-amino acids were purchased from Merck Ltd. (Darmstadt, Germany). Fmoc-NH-SAL-MBHA resin (100–200 mesh); N,N-diisopropylethylamine (DIPEA); and trifluoroacetic acid (TFA) were purchased from Watanabe Chemical Industries Ltd. (Hiroshima, Japan). 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyl uronium hexafluorophosphate (HBTU) and 1-hydroxybenzotriazole (HOBt) were purchased from Kokusan Chemical Co., Ltd. (Tokyo, Japan). Triisopropylsilane (TIS) and xylenol orange were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo, Japan). Hydrochloric acid (HCl); nitric acid; 1,2-ethanedithiol (EDT); CdCl2·2.5H2O; NiCl2·6H2O; MnSO4·5H2O; and tris(hydroxymethyl)aminomethane (Tris) were purchased from Nacalai Tesque Co. Ltd (Kyoto, Japan). ZnCl2 and Multielement Standard Solution W-V were purchased from FUJIFILM Wako Pure Chemical Corporation (Osaka, Japan). Water for the experiments was purified by Milli-Q Integral 3 (Merck Millipore, Darmstadt, Germany). Other solvents and reagents were obtained from commercial suppliers and used without further purification.
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3

Solid-Phase Peptide Synthesis Reagents

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9-Fluorenylmethyloxycarbonyl
(Fmoc) amino acids were purchased from Merck Ltd. (Darmstadt, Germany).
Fmoc-NH-SAL-MBHA resin (100–200 mesh), N,N-diisopropylethylamine (DIPEA), trifluoroacetic acid (TFA),
and (1-cyano-2-ethoxy-2-oxoethylidenaminooxy) dimethylaminomorpholinocarbenium
hexafluorophosphate (COMU) were purchased from Watanabe Chemical Industries
Ltd. (Hiroshima, Japan). 2-(1H-benzotriazole-1-yl)-1,1,3,3-tetramethyl
uronium hexafluorophosphate (HBTU) and 1-hydroxybenzotriazole (HOBt)
were purchased from Kokusan Chemical Co., Ltd. (Tokyo, Japan). 1-Ethyl-3-(3-dimethylaminopropyl)
carbodiimide monohydrochloride (WSCD) was purchased from Peptide Institute,
Inc. (Osaka, Japan). Nitrilotriacetic acid (NTA), nitrilotriacetic
acid disodium salt, 4-dimethylaminopyridine (DMAP), and triisopropylsilane
(TIS) were purchased from Tokyo Chemical Industry Co., Ltd. (Tokyo,
Japan). N,N-diethyldithiocarbamate
(Na-DDTC) for quantification of copper ion was purchased from Nacalai
Tesque Co. Ltd (Kyoto, Japan). Water for experiment was purified by
Milli-Q Integral 3 (Merck Millipore, Darmstadt, Germany). Other solvents
and reagents were also obtained from commercial suppliers and used
without further purification.
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4

Synthesis and Labeling of Affinity Resins

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NovaSyn TGR resin was purchased from Novabiochem (Darmstadt, Germany). Dichloromethane (DCM) and trifluoro acetic acid (TFA) were obtained from Watanabe Chemical Industries (Hiroshima, Japan). N,N-Dimethylformamide (DMF) was obtained from Kanto Chemical (Tokyo, Japan). Chloroacetyl chloride and triisopropylsilane (TIS) were obtained from Tokyo Chemical Industry (Tokyo, Japan). Dimethyl sulfoxide (DMSO) was obtained from Wako Pure Chemical Industries (Osaka, Japan). Methyl-β-cyclodextrin (M-β-CD) was obtained from Sigma Aldrich (St. Louis, MO). NHS-rhodamine (TAMRA SE) and streptavidin protein were obtained from Thermo Fisher Scientific (Waltham, MA).
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