Reagents used in the experiments were obtained from Sigma Aldrich, unless otherwise stated. Protected amino acids were purchased from Trimen Co., (Lodz, Poland). Concentrated solutions of peptides were made in ultrapure water (1 mM) and kept at—20°C until use.
Analytical and semi-preparative RP HPLC was performed using Waters Breeze instrument (Milford, MA, United States) with dual absorbance detector (Waters 2,487) on a Vydac C18 column 5 μm, 4.6 × 250 mm, flow rate 1 mL/min, 50 min linear gradient, and a Vydac C18 column 10 μm, 22 × 250 mm, flow rate 2 mL/min, 20 min linear gradient, respectively, from water/0.1% (v/v) TFA to 80% acetonitrile/20% water/0.1% (v/v) TFA. ESI-MS spectra were obtained on an FTICR (Fourier transform ion cyclotron resonance) Apex-Qe Ultra 7 T mass spectrometer (Bruker Daltonics, Bremen, Germany) equipped with standard ESI source. The instrument was operated in the positive-ion mode and calibrated with the Tunemix™ mixture (Agilent Technologies, Palo Alto, CA, United States). Solutions of peptides were introduced at a flow rate of 3 μL/min.
Analytical and semi-preparative RP HPLC was performed using Waters Breeze instrument (Milford, MA, United States) with dual absorbance detector (Waters 2,487) on a Vydac C18 column 5 μm, 4.6 × 250 mm, flow rate 1 mL/min, 50 min linear gradient, and a Vydac C18 column 10 μm, 22 × 250 mm, flow rate 2 mL/min, 20 min linear gradient, respectively, from water/0.1% (v/v) TFA to 80% acetonitrile/20% water/0.1% (v/v) TFA. ESI-MS spectra were obtained on an FTICR (Fourier transform ion cyclotron resonance) Apex-Qe Ultra 7 T mass spectrometer (Bruker Daltonics, Bremen, Germany) equipped with standard ESI source. The instrument was operated in the positive-ion mode and calibrated with the Tunemix™ mixture (Agilent Technologies, Palo Alto, CA, United States). Solutions of peptides were introduced at a flow rate of 3 μL/min.
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