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MB Sample ID: SA354070
| Local Sample ID: | 230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_01 |
| Subject ID: | SU003379 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003379 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_01 | SA354070 | FL041283 | Pancreatic Cancer cell allografts | Sample source |
| 230221_PE_KPC-Brabletz_1-30-dil_Sample-1-24_01 | SA354070 | FL041283 | epithelial/mixed | Phenotype |
Collection:
| Collection ID: | CO003372 |
| Collection Summary: | Cryo-conserved tumors from subcutaneous allografts described in (Krebs et al. 2017, DOI: 10.1038/ncb3513) were retrieved. |
| Sample Type: | Tumor allograft |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR003388 |
| Treatment Summary: | KPC cells were subcutaneously injected into the flanks of C57BL/6 mice for engraftment as described (Krebs et al. 2017, DOI: 10.1038/ncb3513). Three tumor allografts derived from mesenchymal KPC cell lines (lines KPC550 and KPC701) and epithelial KPC cell lines (lines KPC438 and KPC661) were collected, cryo-conserved and stored at -80°C. |
Sample Preparation:
| Sampleprep ID: | SP003386 |
| Sampleprep Summary: | Phospholipids were extracted from allograft tumor tissue 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 | AN005343 |
|---|---|
| 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: | CH004045 |
| 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: | 5% Water, 95% Acetonitrile; 2 mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005073 |
| Analysis ID: | AN005343 |
| 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 |
