Return to study ST000528 main page
MB Sample ID: SA027778
| Local Sample ID: | POS_Lip_S_24_MCF7 |
| Subject ID: | SU000550 |
| Subject Type: | Cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU000550 |
| Subject Type: | Cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Species Group: | Human |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| POS_Lip_S_24_MCF7 | SA027778 | FL006899 | MCF7 parental | Cell Line |
Collection:
| Collection ID: | CO000544 |
| Collection Summary: | 20 breast cancer cell pellets from four cell lines [MCF7 parental (LSR+), MCF7 Crisper (LSR-), Hs578t parental (LSR-) and Hs578t (LSR++)], with five replicate samples for each cell line were stored at -80˚ until sample preparation. |
| Sample Type: | Cell pellets |
| Storage Conditions: | -80˚C |
Treatment:
| Treatment ID: | TR000564 |
| Treatment Summary: | The cells were cultured under obesogenic media conditions. |
Sample Preparation:
| Sampleprep ID: | SP000557 |
| Sampleprep Summary: | Cell pellets were resuspended in volume of ice cold Lipid Extractions Solvent (50 µg/mL) based on biomass. Contents were transferred to new, pre-labeled 2.0 mL LoBind tube with 10-15 ceramic beads, and homogenized using 2 pulses at 2,000 rpm for 30 sec on a MagNA Lyser. Volume of HPLC-grade water (with 0.02 mg/mL Tryptophan-d5 in water) was added based on biomass. Samples were allowed to sit at room temperature for 10 minutes, then centrifuged at 16,000 rcf for 10 min at 10˚C. For lipidomics, transferred the maximum clean volume of the lower lipid-rich DCM layer to a new, pre-labeled LoBind tube and a 375 µL aliquot was then transferred to a new 1.5 mL, LoBind tube for analysis. An additional 125 µL aliquot of each study sample was transferred to a 7.0 mL glass vial to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (375 µL each), and the remaining was used for 1 Equilibrium sample (column conditioning). Samples were frozen at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in ACN/IPA/H2O (65:30:5 v/v/v), vortexed at 4,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 225 µL of the supernatant was transferred to pre-labeled autosampler vials and 10 µL was injected into OrbiTrap Velos. |
Combined analysis:
| Analysis ID | AN000806 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Waters Acquity |
| Column | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Orbitrap |
| Ion Mode | POSITIVE |
| Units | m/z |
Chromatography:
| Chromatography ID: | CH000581 |
| Chromatography Summary: | Lipidomics |
| Instrument Name: | Waters Acquity |
| Column Name: | Waters Acquity CSH C18 (100 x 2.1mm,1.7um) |
| Column Pressure: | 6000-7500psi |
| Column Temperature: | 50 °C |
| Flow Rate: | 0.25 mL/min |
| Solvent A: | 60% water/40% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Solvent B: | 90% isopropanol/10% acetonitrile; 0.1% formic acid; 10 mM ammonium formate |
| Analytical Time: | 30 min |
| Weak Wash Solvent Name: | 60:40 water/ACN |
| Strong Wash Solvent Name: | 90:10 IPA/ACN |
| Target Sample Temperature: | 8 °C |
| Sample Loop Size: | 20 µL |
| Randomization Order: | Yes |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS000713 |
| Analysis ID: | AN000806 |
| Instrument Name: | Thermo Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 250 °C |
| Collision Energy: | 35 eV |
| Fragmentation Method: | CID |
| Ionization: | ES+ |
| Mass Accuracy: | 10 ppm |
| Source Temperature: | 400 °C |
| Spray Voltage: | 4.0 kV |
| Dataformat: | Profile |
| Resolution Setting: | 30,000 |
| Scan Range Moverz: | 120-2000 m/z |
| Scanning Range: | Normal |
