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MB Sample ID: SA353448
| Local Sample ID: | 190905_AG_Brabletz_ZEB1_EMT_Lipidomics__shGFP_#4_1 |
| 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 |
|---|---|---|---|---|
| 190905_AG_Brabletz_ZEB1_EMT_Lipidomics__shGFP_#4_1 | SA353448 | FL041191 | Breast cancer cell line | Sample source |
| 190905_AG_Brabletz_ZEB1_EMT_Lipidomics__shGFP_#4_1 | SA353448 | 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 |
