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MB Sample ID: SA027802

Local Sample ID:NEG_RP_P_30_JFtotPool3
Subject ID:SU000551
Subject Type:Cells
Subject Species:Homo sapiens
Taxonomy ID:9606
Subject Comments:Cell pellets stored at -80˚C.
Cell Counts:1E 7 cells/pellet
Species Group:Human

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Sample Preparation:

Sampleprep ID:SP000558
Sampleprep Summary:Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For reverse phase, transferred the maximum clean volume of the upper aqueous layer to a new, pre-labeled LoBind tube then a 150 µL aliquot was transferred to a new 1.5 mL, LoBind tube for analysis. An additional 50 µL aliquot of each study sample was combined in a 2 mL LoBind tube to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (150 µL each), and the remainder used for 1 Equilibrium sample (for column conditioning). Samples were placed at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in H2O/Methanol (95:5 v/v), vortexed at 5,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 100 µL of each supernatant was transferred to pre-labeled autosampler vials and 5 µL was injected Synapt G2Si ESI-Q-TOF for data acquisition.
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