The installation of a monomethyl–lysine analog at the mutated cysteine of the H3KC79 (C110A) histone was carried out as described (Simon et al, 2007 (link)) using the 2-chloro-N-methylethanamine hydrochloride (Toronto Research Chemicals C428323) to generate H3KC79me1 (C110A). The installation of the analog was confirmed by electrospray ionization mass spectrometry on a LC-ESI-QTOF Agilent 6538 mass spectrometer and immunoblotting against H3K79me1 (Fig S2 ).
Agilent 6538
Manufactured by Agilent Technologies
Sourced in France, United States
The Agilent 6538 is a high-resolution quadrupole time-of-flight (Q-TOF) mass spectrometer designed for advanced analytical applications. It provides accurate mass measurements and high-quality data for complex sample analysis.
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4 protocols using agilent 6538
1
Histone H3K79 Monomethylation Analog
2
Quantitative Analysis of Atrazine and Metabolites
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Atrazine and metabolites detection and semi quantitative evaluation were performed by LC-HRMS. The HPLC system (Infinity 1290, Agilent Technologies, France) with DAD, was connected to a Q-TOF micro hybrid quadrupole time of flight mass spectrometer (Agilent 6538, Agilent Technologies, France) with electrospray ionization (ESI). HPLC was carried out on a Thermo Hypersyl Gold C18 (USP L1) column (100 × 2.1 mm, 1.9 µm, 175 A), connected to an Agilent Infinity 1290 HPLC at 40 °C. The solvent system was A: 0.1% formic acid in H2O and B: Acetonitrile. The gradient program began with 5% B, held at % for 0.3 min and ramped to 30% B at 1.7 min and to 95% at 3.5 min, held at 95% for 0.5 min, until decreased to initial condition and held at 5 % for 0.5 min. The flow rate was set at 0.600 mL/min. All compounds’ responses were measured in ESI+ and were calibrated externally. The ESI Gas Temp was 350 °C, at electrospray voltage +3800 V. Drying Gaz was set at 10 L/min and Nebuliser was at 30 psi. Fragment voltage was set at 110 V. HRMS spectrum was registered at 5 Hz in the mass range of 50 to 1200 m/z with internal calibration.
3
Phenolic Profiling of Pea Cultivars
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The phenolic compounds of 10 varieties peas were determined by a Waters Acuity ultra-performance liquid chromatography quadrupole time of flight mass spectrometry (UPLC-QTOF-MS) system (Waters, Milford, MA, USA), which equipped with the Agilent Eclipse Plus C18 column (2.1 mm × 50 mm, 1.8 m) and the work settings as follows: the injection volume, 5 µL; column temperature, 35 ℃; flow rate, 0.2 mL/min; mobile phase A, ultrapure water containing 0.1 % (v/v) formic acid; mobile phase B, methanol solution containing 0.1 % (v/v) formic acid. The mass spectrometry performed by a quadrupole time of flight mass spectrometer (Agilent6538), equipped with an electrospray ion source (ESI), and full-scan negative ion mode scanning. The data were obtained in the mass range of 50–1700 m/z. The detector conditions were as follows: capillary voltage, 4.0 kV; sampling cone voltage, 60 V; extraction cone voltage, 40 V; the temperature of dry gas (N2), 350 ℃; the flow rate of the cone gas, 10.0 L/min; atomizing gas pressure, 40 psi; The voltage of cracking, 175 V; ramp collision energy, 20 eV.
4
UHPLC-MS Analysis of Metabolites
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UHPLC-MS analysis was performed on an Agilent 1290 Infinity LC system coupled to an Agilent 6538 accurate-mass quadrupole time-of-flight (Q-TOF) mass spectrometer (Agilent, Palo Alto, CA, USA). Chromatographic separations were performed on an Acquity UHPLC HSS T3 column (2.1 × 100 mm, 2.5 μm, Waters, Milford, MA, USA) at 25 °C, and the injection volume was 3 μL at a flow rate of 0.4 mL/min. The mobile phase consisted of 0.1% formic acid (A) and ACN modified with 0.1% formic acid (B). The gradient program was as follows: 5% B over 0–2 min, 5–95% B over 2–13 min, and 95% B over 13–19 min. The capillary voltage was 4 kV for positive ion mode and 3.5 kV for negative ion mode. The drying gas flow rate was 11 L/min at 350 °C. The nebulizer pressure was 45 psig, the fragmentor voltage was 120 V, and the skimmer voltage was 60 V. Data were collected in a centroid mode and the mass range was m/z 100–1100 using an extended dynamic range.
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