Summary of Study ST001359

This data is available at the NIH Common Fund's National Metabolomics Data Repository (NMDR) website, the Metabolomics Workbench, https://www.metabolomicsworkbench.org, where it has been assigned Project ID PR000929. The data can be accessed directly via it's Project DOI: 10.21228/M8M98M This work is supported by NIH grant, U2C- DK119886.

See: https://www.metabolomicsworkbench.org/about/howtocite.php

This study contains a large results data set and is not available in the mwTab file. It is only available for download via FTP as data file(s) here.

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Study IDST001359
Study TitleMonophasic lipidomics extraction in cancer cell line
Study TypeLipidomics
Study SummaryWe performed a comprehensive characterization of a monophasic extraction method in cancer cell lines. We used pharmacological perturbation on HepG2 cells to identify changes in different lipid families. We optimized the MS parameters, the chromatography and the data analysis to perform rapid and robust lipidomics analysis from cell lines.
Institute
Beatson Institute for Cancer Research
DepartmentMetabolomics
LaboratoryMetabolomics
Last NameRodriguez Blanco
First NameGiovanny
AddressGarscube State, Switchback road, Glasgow
Emailg.blanco@ed.ac.uk
Phone+447526056849
Submit Date2020-03-26
Raw Data AvailableYes
Raw Data File Type(s)raw(Thermo)
Analysis Type DetailLC-MS
Release Date2020-09-29
Release Version1
Giovanny Rodriguez Blanco Giovanny Rodriguez Blanco
https://dx.doi.org/10.21228/M8M98M
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Sample Preparation:

Sampleprep ID:SP001441
Sampleprep 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.
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