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3 protocols using fmoc tyr tbu oh

1

Peptide Synthesis and Characterization

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The conventional amino acids, Fmoc-Ala, Fmoc-Lys(Boc)-OH, Fmoc-Arg(Pbf)-OH, Fmoc-Gly, Fmoc-His(Trt)-OH, Fmoc-Leu, Fmoc-Ser(tBu)-OH, Fmoc-Thr(OtBu)-OH, Fmoc-Trp(Boc)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Asn(Trt)-OH, Fmoc-Ile-OH, Fmoc-Phe-OH, Fmoc-Cys(Trt)-OH, Fmoc-Pro-OH, were acquired from GL Biochem Ltd. (Shanghai, China). N,N′-diisopropylcarbodiimide (DIC), Oxyma pure, 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium 3-oxid hexafluorophosphate (HATU), N,N-Diisopropylethylamine, triisopropylsilane (TIS), 1,1,1,3,3,3-Hexafluoro-2-propanol (HFIP), nonadecanoic acid (C19) were purchased from Sigma-Aldrich (Italy). Rink amide p-methylbenzhydrylamine (MBHA) resin, Fmoc-L-Lys(Mtt)-OH, piperidine, and trifluoroacetic acid (TFA) were purchased from Iris Biotech GmbH (Marktredwitz, Germany). Anhydrous solvents [N,N-dimethylformamide (DMF) and dichloromethane (DCM)], Doxo-EMCH, matrix metalloproteinase-9 (MMP-9) and dialysis tubing benzoylated, Nile Red, and Thioflavin T were purchased from Sigma-Aldrich (Milan, Italy). Silver nitrate (AgNO3), and trisodium citrate were obtained from Sigma-Aldrich (Luzais, Saint-Quentin-Fallavier, France). Ultrapure water was produced using a Barnstead EASYpure RoDi system (Thermo Fisher Scientific, Villebon sur Yvette, France). All chemicals and reagents were of analytical grade and used as received.
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2

Peptide Synthesis and Functionalization

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APTES,
dibenzocyclooctyl-N-hydroxy succinimide (DBCO-NHS), N-(dimethylaminopropyl)-N′-carbodiimide
hydrochloride (EDC), triisopropylsilyl
chloride, and 2-azidoacetic acid were procured from Sigma-Aldrich.
Ethanol (ACS grade), potassium hexacyanoferrate(III), potassium hexacyanoferrate(II),
sodium phosphate dibasic, and sodium phosphate monobasic were purchased
from Merck Chemicals. All chemicals were of analytical reagent (AR)
grade and used without further purification. Aqueous solutions were
prepared using Millipore water received from Milli-Q system (Millipore
Inc.). 9-Fluorenylmethyloxycarbonyl (Fmoc)-protected amino acids (Fmoc-Gln(Trt)-OH,
Fmoc-Asn(Trt)-OH, Fmoc-Gly-OH, Fmoc-Pro-OH, Fmoc-Leu-OH, and Fmoc-Ile-OH)
and Fmoc-Rink-Amide-MBHA resin were procured from Iris Biotech GMPH.
Fmoc-Cys(Trt)-OH and Fmoc-Tyr(tBu)-OH were purchased
from GL Biochem Ltd.
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3

Solid-Phase Peptide Synthesis Protocol

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PL-Wang resin (0.4 mmole/g) was obtained from Varian, Inc. (Shropshire, UK). Fmoc-Ser(tBu)-OH, Fmoc-Met-OH, Fmoc-Leu-OH, Fmoc-Ile-OH, Fmoc-Ala-OH, Fmoc-Lys(Boc)-OH, Fmoc-Asp(OtBu)-OH, Fmoc-Asn(trt)-OH, Fmoc-Glu(OtBu)-OH, Fmoc-Phe-OH, Fmoc-Gln(trt)-OH, Fmoc-Tyr(tBu)-OH, Fmoc-Arg(pbf)-OH, N,N′-diisopropylcarbodiimide (DIC), 4-dimethylaminopyridine (DMAP) and triisopropylsilane (TIS) were obtained from GL Biochem Ltd. (Shanghai, China). Fmoc-Val-OH, Fmoc-Gly-OH, Fmoc-His(trt)-OH and N,N,N′,N′-tetramethyl-uronium-hexafluoro-phosphate (HBTU) were obtained from CS Bio Co. (Menlo Park, USA). Dimethylformamide (DMF), dichloromethane (DCM) and dimethyl sulfoxide (DMSO) were obtained from Daejung Chemicals & Metals Co., Ltd. (Siheung, Korea). Anisole, piperidine and N,N-diisopropylethylamine (DIEA) were obtained from Sigma-Aldrich (St. Louis, USA). Trifluoroacetic acid (TFA) was obtained from Yakuri Pure Chemicals Co., Ltd. (Kyoto, Japan). Ether, Anhydrous was obtained from J.T. Baker (Center Valley, USA). Zoletil® (Virbac) and Rompun® (Bayer Parma) were obtained from SMP animal medicine (Suwon, Korea). Protease inhibitor cocktail was obtained from Roche Diagnostics (IN, USA). EDTA treated BD vacutainer® was obtained from Becton, Dickinson and Company (NJ, USA). Human Aβ42 Ultrasensitive ELISA kit was obtained from Invitrogen (CA, USA).
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