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MB Sample ID: SA205468
Local Sample ID: | 2cCre |
Subject ID: | SU002225 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male and female |
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Combined analysis:
Analysis ID | AN003501 | AN003502 |
---|---|---|
Analysis type | MS | MS |
Chromatography type | Ion Chromatography | None (Direct infusion) |
Chromatography system | ThermoDionexICS3000 | TriVersa NanoMate |
Column | ThermoDionexAS11/AG11 | TriVersa NanoMate |
MS Type | ESI | ESI |
MS instrument type | QTRAP | QTRAP |
MS instrument name | ABI Sciex 3200 QTrap | ABI Sciex 6500 QTrap |
Ion Mode | NEGATIVE | POSITIVE |
Units | mol% | pmol/10E6 cells |
Chromatography:
Chromatography ID: | CH002587 |
Chromatography Summary: | The HPLC-system was controlled by the software Chromeleon VS 6.8 and DCMS-Link VS1.1 (Dionex) in combination with Analyst 1.4.1 (Applied Biosystems). Metabolites were separated on two IonPac AS11HC columns (2 × 250 mm; Dionex) protected by an AG11HC guard column (2 × 50 mm). The elution gradient was generated with water (eluent A) and 100 mm KOH (eluent B) within a total run time of 80 min at a flow rate of 0.25 mL min−1 and a column temperature of 35°C as follows: 0 min, 4%; 0 to 1 min, 4%; 1 to 6 min, 15%; 6 to 12 min, 19%; 12 to 22 min, 20%; 22 to 24 min, 23%; 24 to 27 min, 35%; 27 to 37 min, 38%; 37 to 39 min, 45%; 39 to 44 min, 100%; 44 to 71 min, 100%; 71 to 76 min, 4%; and 76 to 80 min, 4% eluent B. Ref.: Hofmann, J., Bornke, F., Schmiedl, A., Kleine, T., and Sonnewald, U. (2011). Detecting functional groups of Arabidopsis mutants by metabolic profiling and evaluation of pleiotropic responses. 10Front Plant Sci 2, 82. |
Instrument Name: | ThermoDionexICS3000 |
Column Name: | ThermoDionexAS11/AG11 |
Chromatography Type: | Ion Chromatography |
Chromatography ID: | CH002588 |
Chromatography Summary: | Dried lipid extracts were dissolved in 300 μl of methanol. 20 μl of the lipid extract in methanol were loaded into 96-well plates and diluted with 20 μl of 20 mM ammonium acetate in methanol. Lipid infusion and ionization was conducted using Nano-ESI chips with the TriVersa NanoMate operated by the ChipSoft Software (Advion) under the following settings: sample infusion volume: 14 μl, volume of air to aspirate after sample: 1 μl, air gap before chip: enabled, aspiration delay: 0 s, prepiercing: with mandrel, spray sensing: enabled, cooling temperature: 14°C, gas pressure: 0.5 psi, ionization voltage: 1.4 kV, and vent headspace: enabled. Prewetting was done once. Ref.: Kumar, V., Bouameur, J.E., Bar, J., Rice, R.H., Hornig-Do, H.T., Roop, D.R., Schwarz, N., Brodesser, 1120 S., Thiering, S., Leube, R.E., et al. (2015). A keratin scaffold regulates epidermal barrier 1121 formation, mitochondrial lipid composition, and activity. J Cell Biol 211, 1057–1075. Ref.: Kumar, V., Bouameur, J.E., Bar, J., Rice, R.H., Hornig-Do, H.T., Roop, D.R., Schwarz, N., Brodesser, 1120 S., Thiering, S., Leube, R.E., et al. (2015). A keratin scaffold regulates epidermal barrier 1121 formation, mitochondrial lipid composition, and activity. J Cell Biol 211, 1057–1075. |
Instrument Name: | TriVersa NanoMate |
Column Name: | TriVersa NanoMate |
Chromatography Type: | None (Direct infusion) |