Cell pellets were spiked with internal standards (C17-sphingosine Cayman Chemical, Ann Arbor, MI, USA; C17-S1P, Avanti Polar Lipids, Alabaster, AL, USA) and lipids were extracted using a published method (Couttas et al., 2014 (link)). Liquid chromatography tandem-mass spectrometry was performed at PhenoSwitch Bioscience, Inc. (Sherbrooke, QC, Canada). Liquid chromatography gradient was performed using 0.2% v/v formic acid in water and with 0.2% v/v formic acid with 10 mM ammonium formate in methanol on a reversed phase Halo PFP column (Advance Materials Technology, Wilmington, DE, USA), which was maintained at 50°C. Acquisition was performed with an ABSciex TripleTOF 5600 (Sciex, Foster City, CA, USA) equipped with an electrospray interface. Quantification was done using the area under the curve with the MuliQuant software (Sciex). See Supplementary Methods and Supplementary Table 1 for details.
Absciex tripletof 5600
Manufactured by AB Sciex
Sourced in United States
The ABSciex TripleTOF 5600 is a high-resolution mass spectrometry instrument designed for advanced analytical applications. It features a hybrid quadrupole time-of-flight (QTOF) configuration, providing accurate mass measurement and high-resolution data acquisition capabilities.
Automatically generated - may contain errors
2 protocols using absciex tripletof 5600
1
Lipidomic Analysis of Cell Extracts
2
Intact Protein Mass Determination
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The intact mass of SB005 and reference product was determined using LC-ESI-MS (AB SCIEX Triple TOF® 5600+) (SCIEX, Framingham, WA, USA). In total, 10 μg of intact protein sample was injected in time of flight-mass spectroscopy (TOF-MS) through Acquity UPLC (Waters®; Waters Corporation, Milford, MA, USA). The acquired protein spectra were deconvoluted using Bayesian tool of Analyst TF 1.6 software (SCIEX).
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