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2 protocols using 5 ml disposable reaction columns

1

Solid-Phase Peptide Synthesis Protocol

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Tentagel macrobeads were purchased from Rapp Polymere (Germany). All the Fmoc protected amino acids, 2‐(1H‐benzotriazol‐1‐yl)−1,1,3,3‐tetramethyluronium hexafluorophosphate (HBTU), and HOBt were purchased from EMD millipore (Billerica, MA). All primary amines, bromoacetic acid, diisopropylcarbodiimide (DIC), N,N‐diisopropylethylamine (DIPEA), piperidine, trifluoroacetic acid (TFA), and dimethyl formamide (DMF) were purchased from Sigma‐Aldrich (Milwaukee, WI); Dichloromethane (DCM) and acetonitrile were obtained from Honeywell Burdick & Jackson (Morristown, NJ, USA). H‐β‐Ala‐OtBu.HCl was purchased from EMD Biosciences (Gibbstown, NJ, USA). All of the chemical reagents and solvents from commercial sources were used without further purification. 5 mL disposable reaction columns (Intavis AG) were used as reaction vessels for solid‐phase synthesis.
MALDI‐TOF mass spectra were acquired on an Applied Biosystems Voyager‐6115 and Voyager De‐Pro using α‐Cyano‐4‐hydroxycinnamic acid as matrix, while MS/MS (MALDI‐TOF) for sequencing were performed on a 4700 Proteomics Analyzer (Applied Biosystems).
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2

Solid-Phase Synthesis of Peptides

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Tentagel macrobeads were purchased from Rapp Polymere (Germany). All the Fmoc protected amino acids, 2-(1H-benzotriazol-1-yl)-1,1,3,3-tetramethyluronium hexafluorophosphate (HBTU), and HOBt were purchased from EMD millipore (Billerica, MA). All primary amines, bromoacetic acid, diisopropylcarbodiimide (DIC), N,N-diisopropylethylamine (DIPEA), piperidine,, trifluoroacetic acid (TFA), and dimethyl formamide (DMF) were purchased from Sigma-Aldrich (Milwaukee, WI, USA); Dichloromethane (DCM) and acetonitrile were obtained from Honeywell Burdick & Jackson (Morristown, NJ, USA). H-β-Ala-OtBu.HCl was purchased from EMD Biosciences (Gibbstown, NJ, USA). All of the chemical reagents and solvents from commercial sources were used without further purification. 5mL disposable reaction columns (Intavis AG) were used as reaction vessels for solid-phase synthesis.
MALDI-TOF mass spectra were acquired on an Applied Biosystems Voyager-6115 and Voyager De-Pro using α-Cyano-4-hydroxycinnamic acid as matrix, while MS/MS (MALDI- TOF) for sequencing were performed on a 4700 Proteomics Analyzer (Applied Biosystems).
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