Single human H358 cancer cells were isolated by flow cytometry into 96 well plates containing 1 uL of LCMS grade acetonitrile. The cells were placed immediately on dry ice and transported to the lab where they were stored at minus 80C. Dried cell lysate was digested using a solution of 5 nanogram/microliter LCMS grade trypsin (Pierce) in 0.1percent n-Dodecyl-beta-Maltoside Detergent (DDM, Thermo Fisher, 89902) and 50mM TEAB. Two microliters of trypsin solution were used for each cell prior to the plate being tightly sealed with adhesive plate tape (Fisher, 60180-M143) and room temperature overnight digestion. Following digestion, the peptide digest was briefly centrifuged to condense evaporation and the plates were completely dried with vacuum centrifugation. The peptides were resuspended in 3.5 uL of 0.1 percent formic acid, vortexed and centrifuged prior to loading on the autosampler.
[doi:10.25345/C5DB7W19W]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: Single cell proteomics ; diaPASEF ; Buffer modifiers
Principal Investigators: (in alphabetical order) |
Benjamin C Orsburn, Johns Hopkins, USA |
Submitting User: | ben_orsburn |
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