Summary of Study ST002893
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 PR001773. The data can be accessed directly via it's Project DOI: 10.21228/M8J420 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.
Study ID | ST002893 |
Study Title | Folate depletion time-course in MEL cells with analysis for porphyrin metabolites |
Study Summary | Culture of MEL cells in RPMI media containing 100 nM folic acid for 0, 1, 2, 4, 6, or 8 days followed my LC-MS targeting porphyrin metabolites. This is a reverse timecourse where all samples are harvested on the same day. Day 0 in 100 nM folic acid indicates 8 days culture in 2,000 nM folic acid. |
Institute | Boston Children's Hospital, Harvard Medical School |
Department | pathology |
Laboratory | Kanarek Lab |
Last Name | Kanarek |
First Name | Naama |
Address | Enders 1116.2, 300 Longwood Ave, Boston, MA 02115 |
naama.kanarek@childrens.harvard.edu | |
Phone | 6173557433 |
Submit Date | 2023-09-28 |
Num Groups | 5 |
Raw Data Available | Yes |
Raw Data File Type(s) | raw(Thermo) |
Analysis Type Detail | LC-MS |
Release Date | 2024-02-13 |
Release Version | 1 |
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Sample Preparation:
Sampleprep ID: | SP003012 |
Sampleprep Summary: | One million cells from culture were collected via centrifugation, washed with 0.9% NaCl, and resuspended in 150 µl of porphyrin extraction buffer (1:4 ratio of 1.7 M HCl:ACN, 1µM deuteroporphyrin IX (Frontier Scientific, D510-9)) and 0.5 µM isotopically labeled amino acids (Cambridge Isotopes, MSK-A2-1.2)). Samples were vigorously shaken for 20 min at 16ºC in a thermomixer (Eppendorf), sonicated for 10 cycles at 4ºC with 30 sec on and 30 sec off, then incubated at 4ºC for 10 min. Following incubation on ice, samples were centrifuged for 10 minutes at 18,000g to pellet cell debris. The supernatant was collected and 40.5 µl super-saturated MgSO4 and 12 µl 5 M NaCl were added. Samples were vortexed for 30 sec and further shaken for 10 min at 16 ºC in a thermomixer. Finally, a 10 min 10,000 rpm centrifugation was used to separate the organic layer (upper) from the aqueous layer (lower). The upper organic layer was collected. |