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MB Sample ID: SA340746
| Local Sample ID: | m22-L025_SO_03 |
| Subject ID: | SU003264 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 60 |
| Gender: | Female |
| Cell Biosource Or Supplier: | ATCC |
| Cell Strain Details: | BT-474, breast cancer cell line |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003264 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Age Or Age Range: | 60 |
| Gender: | Female |
| Cell Biosource Or Supplier: | ATCC |
| Cell Strain Details: | BT-474, breast cancer cell line |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| m22-L025_SO_03 | SA340746 | FL040302 | BT-474 | Sample source |
| m22-L025_SO_03 | SA340746 | FL040302 | 200 uM | [13C2] Acetate |
| m22-L025_SO_03 | SA340746 | FL040302 | NA | Drug Treatment |
Collection:
| Collection ID: | CO003257 |
| Collection Summary: | One PBS washing, 80% methanol extraction. |
| Collection Protocol Filename: | DM_metabolomics_samples.txt |
| Sample Type: | Cultured cells |
| Collection Method: | 80% methanol extraction |
| Volumeoramount Collected: | 500 ul |
| Storage Conditions: | -80℃ |
| Collection Vials: | 1.5 ml plastic centrifuge tubes |
| Storage Vials: | 1.5 ml plastic centrifuge tubes |
Treatment:
| Treatment ID: | TR003273 |
| Treatment Summary: | Cells were grown in RPMI-1640 supplemented with 200 uM [13C2] acetate and treated with 1 uM Fasnall or 1 uM GSK2194069. |
| Treatment Compound: | Fasnall, GSK2194069 |
| Treatment Dose: | 0 - 1 uM |
| Treatment Doseduration: | 24 h |
| Treatment Vehicle: | DMSO |
| Cell Growth Container: | 6-well plates |
| Cell Media: | RPMI-1640 |
| Cell Envir Cond: | 37C, 5% CO2 |
| Cell Pct Confluence: | ~70% |
Sample Preparation:
| Sampleprep ID: | SP003271 |
| Sampleprep Summary: | Metabolites were extracted with ice-cold 80% methanol. |
| Sampleprep Protocol Filename: | DM_metabolomics_samples.txt |
| Extraction Method: | 80% methanol |
| Extract Enrichment: | None |
| Extract Cleanup: | None |
| Extract Storage: | -80℃ |
| Sample Resuspension: | None |
| Sample Derivatization: | None |
| Sample Spiking: | None |
| Subcellular Location: | Intracellular metabolites |
Combined analysis:
| Analysis ID | AN005163 | AN005164 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | HILIC | HILIC |
| Chromatography system | Thermo Dionex Ultimate 3000 RS | Thermo Dionex Ultimate 3000 RS |
| Column | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF-X Orbitrap | Thermo Q Exactive HF-X Orbitrap |
| Ion Mode | NEGATIVE | POSITIVE |
| Units | AU | AU |
Chromatography:
| Chromatography ID: | CH003908 |
| Instrument Name: | Thermo Dionex Ultimate 3000 RS |
| Column Name: | Merck SeQuant ZIC-pHILIC (150 x 2.1mm,5um) |
| Column Temperature: | 40 |
| Flow Gradient: | A linear solvent gradient of a total duration 22.5 min was starting with 0.2 ml/min flow rate of 80% solvent B, 12.5 min – 30%, 15 min – 30%, 15.2 min – 80%, 20 min – 80%, 21 min – flow rate 0.3 ml/min, 22 min – flow rate 0.3 ml/min, 22.1 min – flow rate 0.2 ml/min. |
| Flow Rate: | 0.2 ml/min |
| Solvent A: | 100% Water; 0.01% ammonium hydroxide; 20 mM ammonium bicarbonate |
| Solvent B: | 100% Acetonitrile |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS004899 |
| Analysis ID: | AN005163 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN. |
| Ion Mode: | NEGATIVE |
| Ion Source Temperature: | 200 |
| Ion Spray Voltage: | 3600 |
| Ionization: | Both |
| Source Temperature: | 200 |
| Automatic Gain Control: | 5e6 |
| Dataformat: | RAW |
| MS ID: | MS004900 |
| Analysis ID: | AN005164 |
| Instrument Name: | Thermo Q Exactive HF-X Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | The HESI ion source voltage was set to the following parameters: 3,600 V in both polarity modes, sheath gas 30, auxiliary gas 5, spare gas 0, probe heater 200 °C, capillary temperature 325 °C, S-Lens RF level 65. The mass spectrometer was set to acquire data in the polarity-switching mode averaging two microscans with 60,000 resolution, automatic gain control (AGC) target 5e6, scan range 72-1080 m/z, and maximum orbitrap injection time (IT) 200 ms. Data were converted into the mzXML format by ProteoWizard and analyzed in MAVEN. |
| Ion Mode: | POSITIVE |
| Ion Source Temperature: | 200 |
| Ion Spray Voltage: | 3600 |
| Ionization: | Both |
| Source Temperature: | 200 |
| Automatic Gain Control: | 5e6 |
| Dataformat: | RAW |
