Samples were analyzed on Agilent 6410B Triple Quadrupole LC/MS system. Peptides were separated on a BioBasic Picofrit C18 capillary column (New Objective). Elution was performed with an acetonitrile gradient from 0 to 100% over 1 h with an overall flow rate of 1 mL/min.
Picofrit c18
Manufactured by New Objective
The Picofrit C18 is a reversed-phase high-performance liquid chromatography (HPLC) column designed for the separation and analysis of a wide range of organic compounds. The column features a silica-based stationary phase with covalently bonded C18 alkyl groups, providing effective retention and separation of both polar and non-polar analytes.
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Lab products found in correlation
2 protocols using picofrit c18
1
Peptide Separation and Analysis by LC/MS
2
Mass Spectrometry Analysis of EVs
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GFP-EVs and Nef/GFP-EVs derived peptides were loaded into the EASY-nLC II (Proxeon Biosystem) coupled to the mass spectrometer LTQ Orbitrap Velos (Thermo Fisher Scientific). The peptides were eluted with a gradient of 2 to 30% acetonitrile in 0.1% formic acid over 120 min at a flow rate of 300 nl/min, using the analytical column PicoFrit C18 (20 cm × 75 μm id, 5 μm; New Objective). The peptides were analyzed by the mass spectrometer operating in the positive mode for data dependent acquisition. Precursor ions (300–1600 m/z) were scanned in the Orbitrap with resolution defined to r = 60.000 and 1.0 × 106 target ions. Up to 20 most intense ions were isolated through a 3 m/z window and activated by collision induced dissociation. Dynamic exclusion was enabled with an exclusion list of up to 500 ions, an exclusion duration of 60 s and a count repetition of 1.
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