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MB Sample ID: SA354067
| Local Sample ID: | 200326_Brabletz_EMT_ZEB1_9_KPC524_3 |
| Subject ID: | SU003378 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003378 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 200326_Brabletz_EMT_ZEB1_9_KPC524_3 | SA354067 | FL041282 | Pancreatic Cancer cells | Sample source |
| 200326_Brabletz_EMT_ZEB1_9_KPC524_3 | SA354067 | FL041282 | mesenchymal/epithelial mixed | Phenotype |
Collection:
| Collection ID: | CO003371 |
| Collection Summary: | Cultured cells were washed, trypsinized, counted and flash-frozen in liquid N2 and stored at -80°C. |
| Sample Type: | Tumor cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003387 |
| Treatment Summary: | Tumor-derived pancreatic cancer KPC cells that are mesenchymal (lines KPC550 and KPC701), mesenchymal/epithelial mixed (lines KPC524 and KPC438), epithelial (lines KPC661 and KPC792) and KPC cell lines with Zeb1 knockout (lines KPCZ519, KPCZ436, KPCZ426) were obtained from tumors of KPC mice as described (Krebs et al. 2017, DOI: 10.1038/ncb3513). |
Sample Preparation:
| Sampleprep ID: | SP003385 |
| 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 | AN005342 |
|---|---|
| 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: | CH004044 |
| 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: | 5% Water, 95% Acetonitrile; 2 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005072 |
| Analysis ID: | AN005342 |
| 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). Phospholipids were analyzed in the negative ion mode, and both fatty acid anion fragments were detected by multiple reaction monitoring (MRM). For quantitation, the mean of both transitions was calculated. For the calculation of relative intensities (i.e., the proportion of lipids), all analyzed signals within the subgroup were summarized (= 100%), and the signals of individual lipid species or lipid subfractions are expressed as percentage of this sum. |
| Ion Mode: | NEGATIVE |
