To confirm the identity of the purified SalB protein as well as monitor conversion from apo to the holo and acylated-holo forms, the protein was digested using the Trypsin Single Proteomics Grade Kit (Sigma Aldrich) and analyzed by High Resolution-LCMS analysis. A 10 μL aliquot was injected onto a Phenomenex Aeris WIDEPORE XB-C18 200 Å, LC column(3.6 μM, 250 mm x 4.6 mm) and analyzed with an Agilent 1260 Infinity LC system coupled to an Agilent 6530 Accurate-Mass Q-TOF. A solvent system of acetonitrile and water both containing 0.1% formic acid (v/v) was used. Peptide fragments were eluted over a 43-min method with a gradient from 5 to 30% acetonitrile over 4 min, 30 to 65% acetonitrile over the next 15 min, and then to 100% over 5 min. 100% acetonitrile was held for 6 minutes before concentration was dropped to 5%. Flow rate was 0.75 mL/min. Eluent was detected using electrospray ionization-mass spectrometry (ESI-MS) monitoring m/z 70–3,200 in positive mode with a speed of 32,500 m/z /s and 30 kV collision energy. MS data were analyzed using Agilent MassHunter Qualitative Analysis B.05.01 software. Peptide fragments were compared to predicted trypsin digest fragments using the ExPASy PeptideMass tool.
Trypsin single proteomics grade kit
Manufactured by Merck Group
The Trypsin Single Proteomics Grade Kit is a laboratory product designed for protein analysis. The kit provides purified trypsin, a widely used protease enzyme, to facilitate the digestion of proteins into smaller peptides for mass spectrometry and other proteomics applications.
Automatically generated - may contain errors
2 protocols using trypsin single proteomics grade kit
1
Mass Spectrometry Identification of Purified SalB
2
Mass Spectrometry Identification of Purified SalB
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To confirm the identity of the purified SalB protein as well as monitor conversion from apo to the holo and acylated-holo forms, the protein was digested using the Trypsin Single Proteomics Grade Kit (Sigma Aldrich) and analyzed by High Resolution-LCMS analysis. A 10 μL aliquot was injected onto a Phenomenex Aeris WIDEPORE XB-C18 200 Å, LC column(3.6 μM, 250 mm x 4.6 mm) and analyzed with an Agilent 1260 Infinity LC system coupled to an Agilent 6530 Accurate-Mass Q-TOF. A solvent system of acetonitrile and water both containing 0.1% formic acid (v/v) was used. Peptide fragments were eluted over a 43-min method with a gradient from 5 to 30% acetonitrile over 4 min, 30 to 65% acetonitrile over the next 15 min, and then to 100% over 5 min. 100% acetonitrile was held for 6 minutes before concentration was dropped to 5%. Flow rate was 0.75 mL/min. Eluent was detected using electrospray ionization-mass spectrometry (ESI-MS) monitoring m/z 70–3,200 in positive mode with a speed of 32,500 m/z /s and 30 kV collision energy. MS data were analyzed using Agilent MassHunter Qualitative Analysis B.05.01 software. Peptide fragments were compared to predicted trypsin digest fragments using the ExPASy PeptideMass tool.
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