(1) Chromatographic conditions: Chromatograph: EASY nLC 1,200 chromatograph (ThermoFisher); Mobile phase: A (water/0.1%FA), B (80%ACN/0.1%FA); Analytical column: 75 μm × 15 cm, Inhouse packed C18 column, 1.9 μm; Flow rate: 0.3 μL/min. (2) Mass spectrometry conditions: Mass spectrometer: Orbitrap Fusion Lumos high-resolution mass spectrometer (ThermoFisher). Software setup parameters: spray voltage, 2.2 kV; parent ion scanning was detected by Orbitrap, mass scanning range: 350–1,500, resolution set to 60 K; daughter ions were detected by Orbitrap, resolution set to 50 K, HCD collision mode, energy 36%.
Easy nlc 1 200 chromatograph
Manufactured by Thermo Fisher Scientific
Sourced in United States
The EASY nLC 1,200 is a capillary high-performance liquid chromatograph (HPLC) designed for sensitive nano-flow separations. It features a flow range of 50 nL/min to 2 μL/min and can be used for a variety of liquid chromatography applications.
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2 protocols using easy nlc 1 200 chromatograph
1
Peptide Identification by Orbitrap Fusion
2
Shotgun Proteomics of Epididymis
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The epididymides were extracted in 50 μL of lysis buffer (8 M urea and 0.2 M ammonium bicarbonate) and sonicated. Each lysate was digested with trypsin and labeled with iTRAQ 4-plex reagent, according to the manufacturer’s instructions. An Orbitrap Q Exactive Plus spectrometer (Thermo Fisher Scientific, Waltham, MA, USA) and an EASY-nLC 1200 chromatograph (Thermo Fisher Scientific, Waltham, MA, USA) were used to generate shotgun proteomics data. Liquid chromatography with tandem mass spectrometry (LC-MS/MS) analysis and database searches were conducted using APRO Science (Tokyo, Japan).
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