Return to study ST001920 main page
MB Sample ID: SA177896
| Local Sample ID: | MDAMB231RR_2_1 |
| Subject ID: | SU001998 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
Select appropriate tab below to view additional metadata details:
Sample Preparation:
| Sampleprep ID: | SP002004 |
| Sampleprep Summary: | Extraction of metabolites and intact lipid species was performed using a modified Folch procedur. Briefly, 1 mL of ice-cold chloroform containing 0.1% BHT and 0.5 mL of ice-cold methanol containing 0.1% BHT were added to the freeze-dried cells and vortexed. The mixture were sonicated for 30 min at 4 °C and incubated for 40 min on ice with shaking. Ice-cold water containing 0.1% BHT (0.38 mL) was added to the mixture for phase separation, and then centrifuged. The upper (methanol) and lower (chloroform) phases were dried under nitrogen gas and used GC-MS and nanoESI-MS analyses, respectively. To conduct metabolic profiling using GC-MS, derivatization reaction was conducted by adding 30 μL of 20,000 μg/mL methoxylamine hydrochloride in pyridine, 10 μL of myristic-d27 acid (300 μg/mL as an internal standard), and 50 μL of BSTFA containing 1% TMCS into dried (methanol phase) sample and incubated for 60 min at 65 °C. After derivatization, 15 μL of each sample was pooled for quality control (QC) and analyzed in sextuplicate. To conduct metabolic profiling using nanoESI-MS, dried (chloroform phase) sample was resuspended with 130 μL of buffer solution (7.5 mM ammonium acetate in methanol-chloroform (9:1, v/v)) and 10 μL of each resuspended sample was pooled for QC and analyzed in sextuplicate. |
| Processing Storage Conditions: | Described in summary |
| Extraction Method: | A modified Folch method |
| Extract Storage: | -80℃ |
| Sample Resuspension: | Described in summary |
| Sample Derivatization: | Described in summary |
