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MB Sample ID: SA273518

Local Sample ID:A375P 8
Subject ID:SU002817
Subject Type:Cultured cells
Subject Species:Homo sapiens
Gender:Female
Cell Biosource Or Supplier:ATCC
Cell Strain Details:A375

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Subject:

Subject ID:SU002817
Subject Type:Cultured cells
Subject Species:Homo sapiens
Gender:Female
Cell Biosource Or Supplier:ATCC
Cell Strain Details:A375

Factors:

Local Sample IDMB Sample IDFactor Level IDLevel ValueFactor Name
A375P 8SA273518FL035348VEHICLETreatment

Collection:

Collection ID:CO002810
Collection Summary:All cell lines had been authenticated in 2021. Cells were expanded to generate enough vials from a single batch before the start of the study. Cell lines were cultured in Dulbecco’s Eagle’s Medium (DMEM) (cat#61965026, Gibco) supplemented with 10% fetal bovine serum (cat#10500064, Gibco) plus 1% penicillin/streptomycin (cat#15140122, Gibco). Cells were grown at 37°C in a 5% CO2 environment.
Sample Type:Melanoma cells

Treatment:

Treatment ID:TR002826
Treatment Summary:Melanoma cells seeded in 6-well plates were treated for 7 days with a high dose (5 or 10μM as indicated) of vemurafenib before switching to a lower concentration of the drug (0,5 or 1μM). Fresh medium and vemurafenib was added once weekly for further 3-4 weeks until arising colonies grew to confluence. Fatty Acid oxidation inhibitors ranolazine, etomoxir or thiroridazine were added once a week.

Sample Preparation:

Sampleprep ID:SP002823
Sampleprep Summary:Metabolites from frozen cell pellets were extracted with cold 5:3:2 MeOH:acetonitrile:water at a concentration of 2 million cells/mL. Samples were vortexed 30 min at 4 degrees C then supernatants clarified by centrifugation (10 min, 10,000 g, 4 degrees C) and transferred to autosampler vials.
Processing Storage Conditions:4℃
Extract Storage:-80℃

Combined analysis:

Analysis ID AN004395 AN004396
Analysis type MS MS
Chromatography type Reversed phase Reversed phase
Chromatography system Thermo Vanquish Thermo Vanquish
Column Phenomenex Kinetex C18 (150 x 2.1mm,1.7um) Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
MS Type ESI ESI
MS instrument type Orbitrap Orbitrap
MS instrument name Thermo Orbitrap Exploris 480 Thermo Orbitrap Exploris 480
Ion Mode NEGATIVE POSITIVE
Units peak area peak area

Chromatography:

Chromatography ID:CH003297
Chromatography Summary:Negative C18
Instrument Name:Thermo Vanquish
Column Name:Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
Column Temperature:45
Flow Gradient:0-0.5 min 0% B, 0.5-1.1 min 0-100% B, 1.1-2.75 min hold at 100% B, 2.75-3 min 100-0% B, 3-5 min hold at 0% B
Flow Rate:450 uL/min
Sample Injection:10 uL
Solvent A:95% water 5% acetonitrile 1 mM ammonium acetate
Solvent B:95% acetonitrile 5% water 1 mM ammonium acetate
Chromatography Type:Reversed phase
  
Chromatography ID:CH003298
Chromatography Summary:Positive C18
Instrument Name:Thermo Vanquish
Column Name:Phenomenex Kinetex C18 (150 x 2.1mm,1.7um)
Column Temperature:45
Flow Gradient:0-0.5 min 5% B, 0.5-1.1 min 5-95% B, 1.1-2.75 min hold at 95% B, 2.75-3 min 95-5% B, 3-5 min hold at 5% B
Flow Rate:450 uL/min
Sample Injection:10 uL
Solvent A:100% water 0.1% formic acid
Solvent B:100% acetonitrile 0.1% formic acid
Chromatography Type:Reversed phase

MS:

MS ID:MS004144
Analysis ID:AN004395
Instrument Name:Thermo Orbitrap Exploris 480
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:We use a Thermo Orbitrap Exploris 120 (not available in the drop down box). Resolution 120,000, scan range 65-975 m/z, maximum injection time 100 ms, microscans 1, automatic gain control (AGC) detection duration 20 msec, source voltage 2.0 kV, capillary temperature 320 C, vaporizer temp 200 C, and sheath gas 50, auxiliary gas 10, and sweep gas 1 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database.
Ion Mode:NEGATIVE
  
MS ID:MS004145
Analysis ID:AN004396
Instrument Name:Thermo Orbitrap Exploris 480
Instrument Type:Orbitrap
MS Type:ESI
MS Comments:We use a Thermo Orbitrap Exploris 120 (not available in the drop down box). Resolution 120,000, scan range 65-975 m/z, maximum injection time 100 ms, microscans 1, automatic gain control (AGC) detection duration 20 msec, source voltage 3.5 kV, capillary temperature 320 C, vaporizer temp 200 C, and sheath gas 50, auxiliary gas 10, and sweep gas 1 (all nitrogen). Data converted to mzXML using RawConverter. Metabolites were annotated and integrated using Maven in conjunction with the KEGG database.
Ion Mode:POSITIVE
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