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MB Sample ID: SA154534
Local Sample ID: | Th2_D4 |
Subject ID: | SU001753 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Not applicable |
Cell Counts: | 5M |
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Sample Preparation:
Sampleprep ID: | SP001759 |
Sampleprep Summary: | The samples were randomized and extracted using a modified version of the previously-published Folch procedure. Briefly, 150 µL of 0.9% NaCl was added to cell pellets, and samples then vortexed and ultrasonicated for 3 minutes. Next, 20 µL of the cell suspension was mixed with 150 µL of the 2.5 µg mL-1 internal standards solution in ice-cold CHCl3:MeOH (2:1, v/v). The internal standard solution contained the following compounds: 1,2-diheptadecanoyl-sn-glycero-3-phosphoethanolamine (PE (17:0/17:0)), N-heptadecanoyl-D-erythro-sphingosylphosphorylcholine (SM(d18:1/17:0)), N-heptadecanoyl-D-erythro-sphingosine (Cer(d18:1/17:0)), 1,2-diheptadeca-noyl-sn-glycero-3-phosphocholine (PC(17:0/17:0)), 1-heptadecanoyl-2-hydroxy-sn-glycero-3-phosphocholine (LPC(17:0)) and 1-palmitoyl-d31-2-oleoyl-sn-glycero-3-phosphocholine (PC(16:0/d31/18:1)). These were purchased from Avanti Polar Lipids, Inc. (Alabaster, AL, USA). In addition, triheptadecanoin (TG(17:0/17:0/17:0)) was purchased from (Larodan AB, (Solna, Sweden). The samples were vortexed and incubated on ice for 30 min after which they were centrifuged at 7800 × g for 5 min. Finally, 60 µL from the lower layer of each sample was collected and mixed with 60 µL of ice cold CHCl3:MeOH (2:1, v/v) in LC vial. The total protein content in cells was measured by the Bradford method. |
Sampleprep Protocol Filename: | parthosen_20210128_003828_PR_CO_Cell_Prep_Protocol.pdf |
Processing Storage Conditions: | -80℃ |
Extract Storage: | -80℃ |