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MB Sample ID: SA162627

Local Sample ID:ovcar8-2
Subject ID:SU001807
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606

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Subject:

Subject ID:SU001807
Subject Type:Cultured cells
Subject Species:Homo sapiens
Taxonomy ID:9606

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
ovcar8-2SA162627FL019213OVCAR8line

Collection:

Collection ID:CO001800
Collection Summary:NCI/ADR-RES (formerly MCF7/Adr) were previously described. NCI/ADR-RES cells were verified to be of ovarian origin by genotyping. OVCAR-8 cells provided by Dr. Ernst Lengyel at the University of Chicago. All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum.
Sample Type:Cultured cells

Treatment:

Treatment ID:TR001820
Treatment Summary:N/A - compared cell lines only

Sample Preparation:

Sampleprep ID:SP001813
Sampleprep Summary:All cancer cell lines were maintained in RPMI 1640 (Sigma R8758) containing glucose (2 mg/ml) and glutamine (0.6 mg/ml) but no pyruvate and was supplemented with 5% Fetal calf serum. Metabolic profile comparisons of OVCAR-8 and NCI/ADR-RES cells were performed on equal cell numbers. Cells were cultured under normal conditions, detached using trypsin-EDTA (0.05 %), counted, normalized to 0.5 x 10e6 in each sample, and cell pellets were snap frozen in liquid nitrogen prior to analysis. Briefly, 2 x 10e6 cells and 20μL cell media were extracted in 1mL and 980μL cold lysis and extraction buffer (methanol : acetonitrile : water, 5:3:2), respectively, and pellets were discarded.

Combined analysis:

Analysis ID AN002815 AN002816
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Phenomenex Kinetex C18 (150 x 2.1mm,2.6um) Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Q Exactive Orbitrap Thermo Q Exactive Orbitrap
Ion Mode POSITIVE NEGATIVE
Units Relative Abundance Relative Abundance

Chromatography:

Chromatography ID:CH002080
Chromatography Summary:Positive Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Positive ion mode - phase A: water, 0.1 % formic acid; B: acetonitrile, 0.1 % formic acid) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described.
Instrument Name:Thermo Vanquish
Column Name:Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
Solvent A:100% water; 0.1 % formic acid
Solvent B:100% acetonitrile, 0.1 % formic acid
Chromatography Type:Reversed phase
  
Chromatography ID:CH002081
Chromatography Summary:Negative Mode 10 μL of water and methanol soluble fractions were run through a Kinetex C18 1.7 μm, 100 x 2.1 mm (Phenomenex) reversed phase column (Negative ion mode – phase A: 1 mM NH4Ac 95:5 water : acetonitrile; phase B: 1 mM NH4Ac 95:5 acetonitrile : water) via an ultra-high performance chromatographic system (UHPLC - Vanquish, Thermo Fisher). UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described.
Instrument Name:Thermo Vanquish
Column Name:Phenomenex Kinetex C18 (150 x 2.1mm,2.6um)
Solvent A:95% water/5% acetonitrile; 1 mM ammonium acetate
Solvent B:95% acetonitrile/5% water; 1 mM ammonium acetate
Chromatography Type:Reversed phase

MS:

MS ID:MS002609
Analysis ID:AN002815
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in positive polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described.
Ion Mode:POSITIVE
  
MS ID:MS002610
Analysis ID:AN002816
Instrument Name:Thermo Q Exactive Orbitrap
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:UHPLC was coupled in line with a high-resolution quadrupole Orbitrap instrument run in negative polarity mode (QExactive, Thermo Fisher) at 70,000 resolution (at 200 m/z). Gradients and other technical parameters and the variants employed herein have been extensively described.
Ion Mode:NEGATIVE
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