This dataset contains 3 data types: trace samples consisting of 2ng and 0.2ng aliquots of HeLa protein digest standard, and single HeLa cells. Pierce™ HeLa protein digest standard and formic acid were purchased from Thermo Fisher Scientific (Waltham, MA). Mobile phase A (0.1% formic acid in water) and mobile phase B (0.1% formic acid in acetonitrile) were respectively prepared from LC-MS grade water and acetonitrile purchased from Honeywell (Charlotte, NC). The digest standard was reconstituted to a final concentration of 200 ng/µL with 100 µL of mobile phase A to form a stock solution. For the experiments, the stock samples were further diluted to 0.2 and 2 ng/µL using the same mobile phase. HeLa cells were cultured from cells purchased from American Type Culture Culture Collection (Manassas, VA). Single HeLa cells were prepared using the nanoPOTS workflow and analyzed by manual LC injection as described previously (29797682) except that cells were isolated into nanowells using the CellenONE platform (Lyon, France) instead of by fluorescence activated cell sorting. Columns: 30-µm-i.d. fused silica capillary columns from Polymicro (Phoenix, AZ) were packed with different materials: Jupiter C18 3.0 µm, 300 Å particles and Kinetex C18 core shell particles of 1.3 µm, 100 Å µm were purchased from Phenomenex (Torrance, CA); BEH C18, 1.7 µm, 130 Å was from Waters (Milford, MA). Column lengths were adjusted to keep the pressure and the linear velocity constant for all columns. The lengths were 50, 9 and 16 cm for Jupiter, Kinetex and BEH columns respectively.
Solid-phase-extraction (SPE) columns were prepared by packing Jupiter C18 particles into 100-µm-i.d. × 5-cm-long fused silica capillaries. The file names contain the sample size and lc packing material.
[doi:10.25345/C5NV69]
[dataset license: CC0 1.0 Universal (CC0 1.0)]
Keywords: single cell
Principal Investigators: (in alphabetical order) |
Ryan Kelly, Brigham Young University, United States |
Submitting User: | hboekweg |
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