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MB Sample ID: SA098896
| Local Sample ID: | VV_18_HEpG2_SDC3 |
| Subject ID: | SU001433 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | HPACC |
| Cell Passage Number: | 10-20 |
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Collection:
| Collection ID: | CO001428 |
| Collection Summary: | Single-phase lipid extraction was performed according to some reported protocols for metabolomics extraction (MacKay et al., 2015). Briefly, media was quickly removed from the plates and each well was washed twice with ice-cold PBS. An extracting solution containing either isopropanol (IPA) or a mixture 1:1 of methanol-butanol (BuMe), kept at 4°C, was used to simultaneously quench all metabolic reactions and to extract intra-cellular lipids. Plates were incubated on dry ice for 20 min and extracted lipids were transferred to 1.5 ml Eppendorf tubes (Stevenage, UK) followed by centrifugation (4°C) at 14000 rpm for 10min to remove the denatured proteins. Supernatants were collected and stored at -80°C prior to LC-MS analysis. Protein pellets attached to the plate were left to dry overnight for the subsequent protein normalisation by the modified-Lowry procedure (MacKay et al., 2015). |
| Collection Protocol Filename: | garodriguezblanco10_20200326_110242_PR_CO_Methods.docx |
| Sample Type: | HepG2 cells |
