Return to study ST001210 main page
MB Sample ID: SA085199
| Local Sample ID: | B5 |
| Subject ID: | SU001277 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Sample Preparation:
| Sampleprep ID: | SP001285 |
| Sampleprep Summary: | Lipid extraction was performed with IPA:H2O (90:10 v/v). Briefly, a 5% (v/v) SPLASH Lipidomix solution was prepared in MeOH and then 50 µL of the diluted solution was added to the cell pellet. Next, 4.95 mL of IPA:H2O (90:10 v/v) were added. Samples were vortexed (30 s), sonicated (2 min) and vortexed again (30 s) to disrupt the pellet. Incubation on ice was continued on a shaker for 1h (500 rpm, 60 min). Samples were centrifuged (3500 rcf, 10 min) and the supernatant was transferred to 15 mL falcon tubes. Samples were dried in an evaporator for 10 hours under nitrogen protection. Dried extract was resuspended in 100 µL of methanol containing odd-chained lipid standards LPC 17:1 and PC 17:0-20:4 at 500 ng/mL and 125 ng/mL, respectively. Sonication (2 min) and vortexing (30 s) were applied to ensure that lipids were not stuck to the surface of the extraction container. Then, samples were centrifuged (3500 rcf, 10 min) and the supernatant was transferred to vials for LC-MS measurements. Quality control (QC) sample was prepared by pooling together 15 µL aliquot of each sample. Samples were measured randomly and QC samples were run at the beginning, at the end and every fifth sample during the sequence. |
| Sampleprep Protocol Filename: | ccalcas_Extraction_and_treatment.pdf |
