Summary of Study ST000154

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 PR000129. The data can be accessed directly via it's Project DOI: 10.21228/M85C7W This work is supported by NIH grant, U2C- DK119886.

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Study IDST000154
Study TitleUse of Aspartate Dehydrogenase by cancer cells
Study SummaryThe human acute lymphoblastic leukemia cell line (Jurkat) was used to isolate a clones based on Noxa expression. JP6 is low Noxa, JP3 is high Noxa. Furthermore, a Noxa expressing plasmid was tranfected and stable clones were selected for a Noxa high model (N5). 10E6 cells were starved of glucose (A) or glutamine (B&C) for 3 hours and then fed 13C 1,2 glucose (A), 15N alpha nitrogen glutamine (B) or 15N amide nitrogen glutamine (C) for 24 hours. Cells were washed 1X in ice cold PBS and resuspended in -20C methanol. Quenched cells were snap frozen and stored at -80. For experiment A we would like to observe the contribution of labeled glucose into the synthesis of amino acids, specifically glycine and serine. For experiment B we are most interested in the nitrogen incorporation into aspartate. SCB edits: This fluxomics study requires measuring amino acids for M, M+1, M+2, and M+3 prioritizing glycine, serine, aspartate, asparagine, ornithine, citrulline, then as many as possible using a diamond hydride column and LCMS method (see CEvans).
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
Email mkachman@umich.edu
Submit Date2015-01-21
Num Groups9
Total Subjects27
Raw Data AvailableYes
Raw Data File Type(s)d
Uploaded File Size3.3 G
Analysis Type DetailGC/LC-MS
Release Date2015-04-24
Release Version1
Maureen Kachman Maureen Kachman
https://dx.doi.org/10.21228/M85C7W
ftp://www.metabolomicsworkbench.org/Studies/ application/zip

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Combined analysis:

Analysis ID AN000245 AN000246
Analysis type MS MS
Chromatography type HILIC GC
Chromatography system Agilent 1260 Agilent 7890A
Column Phenomenex Luna NH2 (150 x 1mm, 3um) Agilent DB5-MS (30m × 0.25mm, 0.25um)
MS Type ESI EI
MS instrument type QTOF Single quadrupole
MS instrument name Agilent 6520 QTOF Agilent 5975C
Ion Mode NEGATIVE POSITIVE
Units Area Area

MS:

MS ID:MS000195
Analysis ID:AN000245
Instrument Name:Agilent 6520 QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
Acquisition Parameters File:QTOF-002-HILIC-35min-1mm.m
Processing Parameters File:EX00383-MassHunterQuant-DataAnalysis-Method.m
  
MS ID:MS000196
Analysis ID:AN000246
Instrument Name:Agilent 5975C
Instrument Type:Single quadrupole
MS Type:EI
Ion Mode:POSITIVE
Acquisition Parameters File:ALPHA_KETO_ACIDS-FULL.M
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