Purified ArtI was alkylated with chloroacetamide or reduced with DTT and alkylated with chloroacetamide. Electrospray mass spectra were recorded on a Bruker micrOTOF-Q II mass spectrometer. An aliquot of each sample, corresponding to approximately 20 picomoles of protein was desalted on-line by reverse-phase HPLC on a Phenomenex Jupiter C4 column (5 µm, 300Ǻ, 2.0 mm x 50 mm) running on an Agilent 1100 HPLC system at a flow rate of 0.2 ml/min using a short water, acetonitrile, 0.05% trifluoroacetic acid gradient. The eluent was monitored at 280 nm and then directed into the electrospray source, operating in positive ion mode, at 4.5 kV and mass spectra recorded from 500-3000 m/z. Data was analysed and deconvoluted to give uncharged protein masses with Bruker's Compass Data Analysis software.
Jupiter c4 column
Manufactured by Agilent Technologies
The Jupiter C4 column is a high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of compounds. It features a C4 stationary phase, which provides excellent retention and resolution for both polar and non-polar analytes. The column is suitable for a variety of applications, including the analysis of proteins, peptides, and other biomolecules.
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3 protocols using jupiter c4 column
1
Protein Mass Spectrometry Analysis
2
Electrospray Mass Spectrometry of Proteins
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Electrospray mass spectra were recorded on a Bruker micrOTOF-Q II mass spectrometer. Samples were desalted on-line by reverse-phase HPLC on a Phenomenex Jupiter C4 column (5 μm, 300 Ǻ, 2.0 mm × 50 mm) running on an Agilent 1100 HPLC system at a flow rate of 0.2 mL/min using a short water, acetonitrile, 0.05% trifluoroacetic acid gradient. The eluant was monitored at 214 nm & 280 nm and then directed into the electrospray source, operating in positive ion mode, at 4.5 kV and mass spectra recorded from 500–3,000 m/z. Data was analysed and deconvoluted to give uncharged protein masses with Bruker’s Compass Data Analysis software.
3
Electrospray Mass Spectrometry Protein Analysis
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The electrospray mass spectrum was recorded on a Bruker micrOTOF‐Q II mass spectrometer. An aliquot of protein in solution, corresponding to approximately 20 pmol of protein, was desalted online by reverse‐phase high performance liquid chromatography (HPLC) on a Phenomenex Jupiter C4 column (5 µm, 300 Ǻ, 2.0 mm × 50 mm) running on an Agilent 1100 HPLC system at a flow rate of 0.2 ml/min using a short water, acetonitrile, 0.05% trifluoroacetic acid gradient. The eluent was monitored at 280 nm and directed into the electrospray source, operating in positive ion mode, at 4.5 kV and mass spectra recorded from 500 to 3,000 m/z. Data were analyzed and deconvoluted to give uncharged protein masses using Bruker's Compass Data Analysis software.
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