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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Project:

Project ID:PR000595
Project DOI:doi: 10.21228/M8RQ4G
Project Title:Purine and TCA Measurements in SS Hypertension
Project Type:MS analysis
Project Summary:Based on RNA-seq ontological and Bayesian analyses, both purine levels and TCA-cycle intermediates have been nominated as being regulated by pressure in the SS rat. This project seeks to verify the previous RNA-seq prediction.
Institute:Medical College of Wisconsin
Department:Physiology
Laboratory:Cowley Lab
Last Name:Cowley
First Name:Allen
Address:Milwaukee, WI
Email:cowley@mcw.edu
Phone:414-955-8277

Subject:

Subject ID:SU000862
Subject Type:MOUSE
Subject Species:Mus musculus
Taxonomy ID:10090
Species Group:Mammal

Factors:

Subject type: MOUSE; Subject species: Mus musculus (Factor headings shown in green)

mb_sample_id local_sample_id Sample type
SA046609S00026317Cell Line | Treatment:DUB inhibitor | Type:RAW
SA046610S00026318Cell Line | Treatment:DUB inhibitor | Type:RAW
SA046611S00026319Cell Line | Treatment:DUB inhibitor | Type:RAW
SA046612S00026315Cell Line | Treatment:DUB inhibitor | Type:RAW
SA046613S00026316Cell Line | Treatment:DUB inhibitor | Type:RAW
SA046614S00026311Cell Line | Treatment:Vehicle | Type:RAW
SA046615S00026310Cell Line | Treatment:Vehicle | Type:RAW
SA046616S00026312Cell Line | Treatment:Vehicle | Type:RAW
SA046617S00026313Cell Line | Treatment:Vehicle | Type:RAW
SA046618S00026314Cell Line | Treatment:Vehicle | Type:RAW
SA046619S00026320Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM
SA046620S00026321Macrophages | Treatment:Control-Other_Cells | Type:Primary BL6 BMM
Showing results 1 to 12 of 12

Collection:

Collection ID:CO000856
Collection Summary:-
Sample Type:Cultured cells

Treatment:

Treatment ID:TR000876
Treatment Summary:-

Sample Preparation:

Sampleprep ID:SP000869
Sampleprep Summary:-
Sampleprep Protocol Filename:A043-TCA-Plus-20170713.pdf

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

MS:

MS ID:MS001228
Analysis ID:AN001336
Instrument Name:Agilent 6520B QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
Analysis Protocol File:A043-TCA-Plus-20170713.pdf
  
MS ID:MS001229
Analysis ID:AN001337
Instrument Name:Agilent 6520B QTOF
Instrument Type:QTOF
MS Type:ESI
Ion Mode:NEGATIVE
Analysis Protocol File:A043-TCA-Plus-20170713.pdf
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