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MB Sample ID: SA242388
| Local Sample ID: | 20220820_tufts_MAMfrac_pos_rep8_KO_4ul |
| Subject ID: | SU002511 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | HEK293T |
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Combined analysis:
| Analysis ID | AN003944 | AN003945 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um) | Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive Plus Orbitrap | Thermo Q Exactive Plus Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | peak area | peak area |
Chromatography:
| Chromatography ID: | CH002920 |
| Chromatography Summary: | Lipids were identified and quantitatively measured using ultra high-performance liquid-chromatography high-resolution tandem MS/MS (UHPLC-MS/MS) as recently described89,90. Separation of lipids was done by reverse-phase chromatography using a Waters ACQUITY CSH C18 column (150x2.1mm; 1.7um) at 60°C using a Vanquish UHPLC system (Thermo Scientific) and two solvents: solvent A (40:60 water-methanol plus 10mM ammonium formate and 0.1% formic acid) and solvent B (10:90 methanol-isopropanol plus 10mM ammonium formate and 0.1% formic acid). UHPLC was performed at a 0.25 ml per min flow rate for 30 min per sample, starting at 25% solvent B and ending at 100% solvent B as described. The column was washed and equilibrated between samples. Samples were run in a semi-random order where WT or UBXD8 KO samples were interspersed with blank samples. Each sample was repeatedly analyzed using various injection volumes (4, 8, 16ul for positive mode and 5, 10, and 15ul for negative mode). |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters ACQUITY UPLC CSH C18 (150 x 2.1mm,1.7um) |
| Column Temperature: | 60 |
| Flow Gradient: | 25% to 100% |
| Flow Rate: | 0.25mL per min |
| Solvent A: | 40% water; 60% methanol 10mM ammonium formate and 0.1% formic acid |
| Solvent B: | 10% methanol; 90% isopropanol 10mM ammonium formate and 0.1% formic acid |
| Chromatography Type: | Reversed phase |
