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MB Sample ID: SA358649
| Local Sample ID: | sgLuc -Gln 60 2 |
| Subject ID: | SU003427 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Female |
| Cell Strain Details: | MCF7 |
| Cell Primary Immortalized: | Immortalized |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003427 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Gender: | Female |
| Cell Strain Details: | MCF7 |
| Cell Primary Immortalized: | Immortalized |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| sgLuc -Gln 60 2 | SA358649 | FL041868 | Human breast cancer cells | Sample source |
| sgLuc -Gln 60 2 | SA358649 | FL041868 | Ctrl | Genotype |
| sgLuc -Gln 60 2 | SA358649 | FL041868 | (-Gln)1hr | Treatment |
Collection:
| Collection ID: | CO003420 |
| Collection Summary: | Cells were washed with PBS and fed media containing either complete serine (285uM) or low serine (50uM) for six hours. Cells were washed with saline and harvested in cold GCMS-grade methanol with norvaline diluted in water. Polar metabolites were extracted with the addition of chloroform. Samples were dried under constant air flow for 2 hours. |
| Collection Protocol Filename: | CL_PM2024.pdf |
| Sample Type: | Cultured cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003436 |
| Treatment Summary: | Cells are either control (sgLuc) or ASCT2-KO (sgSLC1A5). All groups were plated in triplicate in a six well plate 48hours prior to treatment. Cells were treated with either complete media, or media containing low serine for six hours, or media lacking glutamine (-Q) for one hour prior to harvesting. |
| Treatment Doseduration: | 1-6 hours |
| Cell Growth Container: | Corning 6-well plates |
| Cell Media: | RPMI w/o Glucose, Sodium Pyruvate, Amino acids |
| Cell Pct Confluence: | 80% |
Sample Preparation:
| Sampleprep ID: | SP003434 |
| Sampleprep Summary: | Samples were harvested in cold GCMS-grade methanol with norvaline diluted in water. Polar metabolites were separated with chloroform and air dried for two hours. Samples were then rehydrated in 15uL MOX reagent and heated at 37C for 90 minutes. Then 20uL of TBDMS was added to each sample and heated at 60C for 60 minutes. |
| Sampleprep Protocol Filename: | CL_PM2024.pdf |
Combined analysis:
| Analysis ID | AN005417 |
|---|---|
| Analysis type | MS |
| Chromatography type | GC |
| Chromatography system | Agilent 7890B |
| Column | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| MS Type | EI |
| MS instrument type | Single quadrupole |
| MS instrument name | Agilent 5977B |
| Ion Mode | POSITIVE |
| Units | Abundance |
Chromatography:
| Chromatography ID: | CH004107 |
| Chromatography Summary: | All samples were analyzed by GC/MS using a HP-5MS Ultra Inert GC column (19091S-433UI, Agilent Technologies) installed in an Agilent 7890B gas chromatograph coupled to an Agilent 5779B mass spectrometer. Helium was used as the carrier gas. One microliter was injected (split inlet) at 280 degrees C. After injection, the GC oven was held at 60 degrees C for 1 minute before ramping to 320 degrees C at 10C/min and held for 9 minutes at the maximum temperature. |
| Instrument Name: | Agilent 7890B |
| Column Name: | Agilent HP5-MS (30m x 0.25mm, 0.25 um) |
| Column Temperature: | 60-320 |
| Flow Gradient: | N/A |
| Flow Rate: | 1.5mL/min |
| Solvent A: | N/A |
| Solvent B: | N/A |
| Chromatography Type: | GC |
MS:
| MS ID: | MS005143 |
| Analysis ID: | AN005417 |
| Instrument Name: | Agilent 5977B |
| Instrument Type: | Single quadrupole |
| MS Type: | EI |
| MS Comments: | The MS system operated under electron impact ionization mode at 70 eV and the MS source and quadrupole were held at 230 degrees C and 150 degrees C respectively. Peak areas were determined using MassHunter software. |
| Ion Mode: | POSITIVE |
| Analysis Protocol File: | CL_GCMS2024.pdf |
