AMA (100 μM) was incubated with an equimolar concentration of ZnSO4 for 10 min at room temperature. The mixture was analyzed with ESI–MS by direct infusion using a Thermo Scientific LTQ OrbiTrap XL ETD Hybrid Ion Trap-Orbitrap Mass Spectrometer (Thermo Fisher Scientific) at a flow rate of 5 μl⋅min−1 with a spray voltage of 4.5 kV and capillary temperature of 350 °C. The instrument was operated in Fourier transform and positive-ion modes with MS scans between 100 and 2000 m/z. Mass spectra were analyzed using Mmass—open source MS tool.
Ltq orbitrap xl etd hybrid ion trap orbitrap mass spectrometer
Manufactured by Thermo Fisher Scientific
Sourced in Japan
The LTQ OrbiTrap XL ETD Hybrid Ion Trap-Orbitrap Mass Spectrometer is a high-performance analytical instrument designed for advanced mass spectrometry applications. It combines the capabilities of an ion trap and an Orbitrap mass analyzer, providing high mass resolution, accurate mass measurement, and tandem mass spectrometry (MS/MS) capabilities.
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2 protocols using ltq orbitrap xl etd hybrid ion trap orbitrap mass spectrometer
1
Analysis of AMA-Zn Complexation by ESI-MS
2
Proteomic Profiling of Purified Enzymes
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Sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) of the purified enzymes was performed with SYPRO Ruby Protein Gel Stain (Thermo Fisher Scientific). The protein bands were excised and digested with trypsin (Promega, Madison, WI) in a MultiScreen 96-well filter plate (Merck Millipore, Darmstadt, Germany). The tryptic digests were analyzed with nano-LC (Shimadzu, Kyoto, Japan) and an LTQ OrbitrapXL ETD Hybrid Ion Trap-Orbitrap Mass Spectrometer (Thermo Fisher Scientific) and identified using the Mascot database search system (Matrix Science, Boston, MA) and the SProt database. Data processing was performed by Proteome discoverer software version 1.4 (Thermo Fisher Scientific). The precursor mass tolerance was set at 20 ppm, the fragment ion mass tolerance was 0.2 Da, and the false discovery rate was 0.01.
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