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MB Sample ID: SA027793
Local Sample ID: | NEG_RP_S_38_LSRH-L6 |
Subject ID: | SU000551 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Subject Comments: | Cell pellets stored at -80˚C. |
Cell Counts: | 1E 7 cells/pellet |
Species Group: | Human |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU000551 |
Subject Type: | Cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Subject Comments: | Cell pellets stored at -80˚C. |
Cell Counts: | 1E 7 cells/pellet |
Species Group: | Human |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
NEG_RP_S_38_LSRH-L6 | SA027793 | FL006902 | Hs578t+ | Cell Line |
Collection:
Collection ID: | CO000545 |
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: | TR000565 |
Treatment Summary: | Cultured in an obesogenic media. |
Sample Preparation:
Sampleprep ID: | SP000558 |
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 reverse phase, transferred the maximum clean volume of the upper aqueous layer to a new, pre-labeled LoBind tube then a 150 µL aliquot was transferred to a new 1.5 mL, LoBind tube for analysis. An additional 50 µL aliquot of each study sample was combined in a 2 mL LoBind tube to make a QC pool, which was vortexed for 30 sec and aliquoted into 5 Total Pool samples (150 µL each), and the remainder used for 1 Equilibrium sample (for column conditioning). Samples were placed at -80˚C for 60 mins and lyophilized to dryness. Samples were reconstituted in H2O/Methanol (95:5 v/v), vortexed at 5,000 rpm for 2 mins and centrifuged at 16,000 rcf for 4 min. 100 µL of each supernatant was transferred to pre-labeled autosampler vials and 5 µL was injected Synapt G2Si ESI-Q-TOF for data acquisition. |
Combined analysis:
Analysis ID | AN000807 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity |
Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
MS Type | ESI |
MS instrument type | QTOF |
MS instrument name | Waters Synapt G2 Si QTOF |
Ion Mode | NEGATIVE |
Units | m/z |
Chromatography:
Chromatography ID: | CH000582 |
Chromatography Summary: | Reversed-Phase Gradient Seperation |
Methods ID: | RTI-RCMRC-RP |
Methods Filename: | RTI-RCMRC-RP |
Instrument Name: | Waters Acquity |
Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
Column Temperature: | 50 °C |
Flow Rate: | 0.4 mL/min |
Internal Standard: | L-Tryptophan-d5 |
Solvent A: | 100% water; 0.1% formic acid |
Solvent B: | 100% methanol; 0.1% formic acid |
Analytical Time: | 22 min |
Weak Wash Solvent Name: | 10% MeOH |
Weak Wash Volume: | 1000 µL |
Strong Wash Solvent Name: | 80% MeOH |
Strong Wash Volume: | 1000 µL |
Target Sample Temperature: | 8 °C |
Sample Loop Size: | 10 µL |
Sample Syringe Size: | 100 µL |
Randomization Order: | Yes |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS000714 |
Analysis ID: | AN000807 |
Instrument Name: | Waters Synapt G2 Si QTOF |
Instrument Type: | QTOF |
MS Type: | ESI |
Ion Mode: | NEGATIVE |
Capillary Temperature: | 110 °C |
Capillary Voltage: | 1.0 kV |
Collision Energy: | 4 |
Fragmentation Method: | CID |
Helium Flow: | 180 |
Ionization: | ES- |
Ionization Potential: | 5 ppm |
Source Temperature: | 100 °C |
Dataformat: | Continuum |
Desolvation Gas Flow: | 400 L/Hr |
Desolvation Temperature: | 400 °C |
Resolution Setting: | 18000 |
Scan Range Moverz: | 50-1000 m/z |
Scanning Cycle: | 1.0 s |
Tube Lens Voltage: | 74 |