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MB Sample ID: SA353437
Local Sample ID: | 220408_MDA shZEB1_1_10_PE_7 |
Subject ID: | SU003364 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU003364 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
220408_MDA shZEB1_1_10_PE_7 | SA353437 | FL041191 | Breast cancer cell line | Sample source |
220408_MDA shZEB1_1_10_PE_7 | SA353437 | FL041191 | control for Zeb1 stable knockdown | Genotype |
Collection:
Collection ID: | CO003357 |
Collection Summary: | Cultured cells were washed, trypsinized, counted and flash-frozen in liquid N2 and stored at -80°C. |
Sample Type: | Breast cancer cells |
Storage Conditions: | -80℃ |
Treatment:
Treatment ID: | TR003373 |
Treatment Summary: | Human breast cancer MDA-MB-231 wildtype (WT) cells and the stably transduced MDA-MB-231 shZeb1 (stable Zeb1 knockdown) and shCtrl cell lines (control cell line for the stable Zeb1 knockdown) (Spaderna et al. 2008, DOI: 10.1158/0008-5472.CAN-07-5682) were collected, snap-frozen and stored at -80°C. |
Sample Preparation:
Sampleprep ID: | SP003371 |
Sampleprep Summary: | Phospholipids were extracted from cell pellets by successive addition of PBS pH 7.4, methanol, chloroform, and saline to a final ratio of 14:34:35:17. Evaporation of the organic layer yielded a lipid film that was dissolved in methanol and subjected to UPLC-MS/MS. |
Extract Storage: | -80℃ |
Combined analysis:
Analysis ID | AN005315 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Waters Acquity H-Class |
Column | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) |
MS Type | ESI |
MS instrument type | QTRAP |
MS instrument name | ABI Sciex 6500+ |
Ion Mode | NEGATIVE |
Units | relative intensities |
Chromatography:
Chromatography ID: | CH004019 |
Chromatography Summary: | Chromatographic separation of phospholipids was carried out on an Acquity BEH C8 column (1.7 μm, 2.1×100 mm, Waters, Milford, MA) using an Acquity UHPLC. |
Instrument Name: | Waters Acquity H-Class |
Column Name: | Waters ACQUITY UPLC BEH C8 (100 x 2.1mm,1.7um) |
Column Temperature: | 45 |
Flow Gradient: | The gradient was ramped from 75 to 85% B over 5 min and further increased to 100% B within 2 min, followed by isocratic elution for another 2 min |
Flow Rate: | 0.75 mL/min |
Solvent A: | 90% Water, 10% acetonitrile; 2 mM ammonium acetate |
Solvent B: | 95% Acetonitrile, 5% Water; 2 mM ammonium acetate |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS005045 |
Analysis ID: | AN005315 |
Instrument Name: | ABI Sciex 6500+ |
Instrument Type: | QTRAP |
MS Type: | ESI |
MS Comments: | Targeted MRM with pre-optimized settings and subsequent automated integration of selected signals using Analyst 1.6.3 or Analyst 1.7.1 (Sciex). |
Ion Mode: | NEGATIVE |