Return to study ST000528 main page
MB Sample ID: SA027764
Local Sample ID: | POS_Lip_S_37_LSRH-L4 |
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_37_LSRH-L4 | SA027764 | FL006897 | Hs578t+ | 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 |