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4 protocols using amino acids

1

Solid-Phase Peptide Synthesis Workflow

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All reagents and solvents were purchased from commercial sources. Amino acids and 2-chlorotrityl-chloride (CTC) resin were obtained from (Iris Biotech, Marktredwitz, Germany) and N,N-Diisopropylethylamine (DIEA) from (Sigma-Aldrich, St. Louis, MO, USA). N,N,N′,N′-tetramethyl-O-(1H-benzotriazol-1-yl)uronium hexafluorophosphate (HBTU) and 1-cyano-2-ethoxy-2-oxoethyliden(aminooxy) dimethylamino-morpholino-carbenium hexafluoro phosphate (COMU) were a gift from Luxembourg Technologies. Organic solvents were supplied by (Merck, Darmstadt, Germany).
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2

Peptide-receptor binding assay protocol

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Unless otherwise specified, reagents were obtained from commercial sources. LC-MS grade water was purchased from Huberlab (Aesch, Switzerland); methanol and acetonitrile (both HPLC gradient grade) were purchased from VWR (Darmstadt, Germany). Ammonium formate and formic acid were purchased from Sigma-Aldrich (Buchs, Switzerland). Other solvents and reagents were purchased from Merck (Darmstadt, Germany). Fmoc-Arg(Pbf) Wang resin was obtained from Activotec (Cambridge, UK). Amino acids and coupling reagents were purchased from Iris Biotech (Marktredwitz, Germany). Recombinant human receptors and biotinylated human VEGF-A165 were purchased from R&D Systems (Minneapolis, MN, USA). Chemiluminescent, streptavidin-horseradish peroxidase conjugate and DPBS were obtained from Thermo Scientific (Waltham, MA, USA).
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3

Peptide Synthesis Using Resin, Amino Acids

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Resin, amino acids and activators were purchased from Iris Biotech GmbH (Marktredwitz, Germany). N-ethyldiisopropylamine (DIPEA) and bromophenol blue (BPB) were purchased from Merck KGaA (Darmstadt, Germany).
All the abbreviations used can be found in Abbreviation.
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4

Peptide Synthesis and NRP-1 Binding

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Unless otherwise specified, reagents were obtained from commercial sources and used without further purification. Fmoc-Arg(Pbf) Wang resin was purchased from Activotec (Cambridge, UK). Amino acids and coupling reagents were obtained from Iris Biotech (Marktredwitz, Germany). Solvents and reagents were purchased from Merck (Darmstadt, Germany). Recombinant human NRP-1 and biotinylated human VEGF-A165 were purchased from R&D Systems (Minneapolis, MN, USA). Chemiluminescent substrate was purchased from Thermo Scientific (Waltham, MA, USA). Luminescence was measured using a Tecan Infinite F200Pro microplate reader (Männedorf, Switzerland).
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