For size exclusion chromatography, proteins were first buffer-exchanged into 300 mM ammonium acetate, pH 6.8 with Micro Bio-Spin™ 6 columns (Bio-Rad). 100 pmol protein was injected onto a bioZen™ 1.8 μm SEC-2 column (150 x 4.6 mm) equipped with a security guard column (Phenomenex), and eluted with 300 mM ammonium acetate at a flow rate of 0.2 mL min−1 using an UltiMate™ 3000 RSLC (Thermo Fisher Scientific) coupled to an Exactive™ Plus EMR Orbitrap™ mass spectrometer (Thermo Fisher Scientific) equipped with an ESI source and modified to incorporate a quadrupole mass filter and allow for surface-induced dissociation [19 (link)]. Mass spectra were recorded from 1000 - 10000 m/z at 8750 resolution as defined at 200 m/z. The ion injection time was set to 200 ms. Voltages applied to the transfer optics were optimized to allow for optimal ion transmission. Total ion chromatograms were smoothed (Boxcar; 15 points) and overlaid. Mass spectra were deconvoluted and plotted with UniDec version 4.0.0 beta [20 (link)] using the preset high-resolution native parameters adjusted to sample mass every 1 Da.
Exactive plus emr orbitrap mass spectrometer
Manufactured by Thermo Fisher Scientific
The Exactive Plus EMR Orbitrap Mass Spectrometer is a high-resolution mass spectrometer designed for accurate mass measurements. It utilizes Orbitrap technology to provide precise mass analysis of a wide range of compounds.
Automatically generated - may contain errors
Lab products found in correlation
Security guard column, by Phenomenex (1 mentions)
Micro bio spin 6 column, by Bio-Rad (1 mentions)
Ultimate 3000 rslc, by Thermo Fisher Scientific (1 mentions)
Platinum coated emitter, by Thermo Fisher Scientific (1 mentions)
Flash 2000, by Thermo Fisher Scientific (1 mentions)
Icap 6300, by Thermo Fisher Scientific (1 mentions)
5 protocols using exactive plus emr orbitrap mass spectrometer
1
Native Mass Spectrometry of Proteins
2
Native Mass Spectrometry of Protein Complexes
3
Mass Spectrometry Analysis of Protein-Phospholipid-TMAO Interactions
4
High-Resolution Mass Spectrometry Protocol
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High-resolution mass spectra were obtained on a Thermo Scientific Exactive Plus EMR Orbitrap Mass Spectrometer (ThermoFisher Scientific). The electrospray ionization source was operated in the positive mode with spray voltage of 2.5 kV and ion transfer tube temperature at 270 °C. Gas flow was set to defaults for the used 10-µL/min injection which was performed using the syringe pump. Each scan consisted of three microscans with a detection range set over a mass range of 150 to 2,000 m/z.
5
Synthesis and Characterization of U(IV) Complexes
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General information This journal is © The Royal Society of Chemistry 20xx
Please do not adjust margins Please do not adjust margins Unless stated otherwise, all reagents and solvents were purchased from commercial sources and used without further purification.
For U IV reactions, all manipulations were carried out using standard Schlenk and glove box techniques under an atmosphere of dry argon. Solvents were dried by refluxing over potassium and degassed before use. All solvents were stored over potassium mirrors. Most solid reagents were dried under vacuum for four hours and most liquid reagents were dried over 4 Å molecular sieves and distilled before use. Analytical data were obtained by the microanalysis laboratory at the University of Manchester-carbon, hydrogen, nitrogen analysis (CHN) by a Flash 2000 elemental analyser and metals analysis by Thermo iCap 6300 inductively coupled plasma optical emission spectroscopy (ICP-OES).
Thermo Fisher Scientific "Exactive Plus EMR Orbitrap" mass spectrometer was used for mass spectrometry analysis. The samples were ionised in the electrospray ion source operated in both positive and negative modes, ~3.5 kV was applied to the capillary. The ions were detected using electrostatic trap with mass resolution of ~140000 and stability sufficient to achieve a precision of <5 ppm for mass measurements.
Please do not adjust margins Please do not adjust margins Unless stated otherwise, all reagents and solvents were purchased from commercial sources and used without further purification.
For U IV reactions, all manipulations were carried out using standard Schlenk and glove box techniques under an atmosphere of dry argon. Solvents were dried by refluxing over potassium and degassed before use. All solvents were stored over potassium mirrors. Most solid reagents were dried under vacuum for four hours and most liquid reagents were dried over 4 Å molecular sieves and distilled before use. Analytical data were obtained by the microanalysis laboratory at the University of Manchester-carbon, hydrogen, nitrogen analysis (CHN) by a Flash 2000 elemental analyser and metals analysis by Thermo iCap 6300 inductively coupled plasma optical emission spectroscopy (ICP-OES).
Thermo Fisher Scientific "Exactive Plus EMR Orbitrap" mass spectrometer was used for mass spectrometry analysis. The samples were ionised in the electrospray ion source operated in both positive and negative modes, ~3.5 kV was applied to the capillary. The ions were detected using electrostatic trap with mass resolution of ~140000 and stability sufficient to achieve a precision of <5 ppm for mass measurements.
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