Summary of Study ST000835

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 PR000595. The data can be accessed directly via it's Project DOI: 10.21228/M8RQ4G This work is supported by NIH grant, U2C- DK119886.

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Study IDST000835
Study TitleDeubiquitinase inhibitor compound C6 effect on cellular metabolism
Study TypeMS analysis
Study Summary6.0 X 10^6 cells were plated in 10 cm Tissue Culture dishes and allowed to recover overnight at 37 degrees and 5% CO2. In the am, supernatant was removed, and 12 mL of fresh medium with either 2.5 micromolar Compound C6 or a v/v match of DMSO (vehicle control) was added to the cells. Cells were incubated at 37 degrees in 5% CO2 for 30 minutes. Cells were washed 3X with cold DPBS++ and 10 mL of medium was added to the cells with either 1.0 micromolar Compound C6 or v/v match of DMSO. Cells were incubated at 37 degrees and 5% CO2 for 7.0 hours. Cells were washed with 10 mL of 150 mM Ammonium Acetate, swirled, washed again with 5 mL, and immediately quenched with liquid nitrogen. Cells were then frozen at -80 degrees.
Institute
University of Michigan
DepartmentBiomedical Research Core Facilities
LaboratoryMetabolomics core
Last NameKachman
First NameMaureen
Address6300 Brehm Tower, 1000 Wall Street, Ann Arbor, MI 48105-5714
Emailmkachman@med.umich.edu
Phone(734) 232-8175
Submit Date2017-08-02
Num Groups7
Total Subjects1
Study CommentsI request that the metabolites of the TCA+ assay that are to be assessed quantitatively please be sure to include ATP, ADP, AMP, NAD, NADH, NADP, NADPH, E4P, S7P, 6PG, G3P, H6P, and Lactate in particular. Also, I would request that both reduced and oxidized Glutathione as well as Fructose 1,6-bisphosphate be included in the non-quantitative data.
Raw Data AvailableYes
Raw Data File Type(s)d
Analysis Type DetailLC-MS
Release Date2018-02-07
Release Version1
Maureen Kachman Maureen Kachman
https://dx.doi.org/10.21228/M8RQ4G
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN001336 AN001337
Analysis type MS MS
Chromatography type HILIC HILIC
Chromatography system Agilent Agilent
Column Phenomenex Luna NH2 (150 x 1mm,3um) Phenomenex Luna NH2 (150 x 1mm,3um)
MS Type ESI ESI
MS instrument type QTOF QTOF
MS instrument name Agilent 6520B QTOF Agilent 6520B QTOF
Ion Mode NEGATIVE NEGATIVE
Units ISTD response ratio pmol/ug protein

Chromatography:

Chromatography ID:CH000934
Instrument Name:Agilent
Column Name:Phenomenex Luna NH2 (150 x 1mm,3um)
Solvent A:100% water; 5 mM ammonium acetate, pH 9.9
Solvent B:100% acetonitrile
Chromatography Type:HILIC
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