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MB Sample ID: SA022561
| Local Sample ID: | TB231TDM48R3N |
| Subject ID: | SU000463 |
| Subject Type: | Cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Biosource Or Supplier: | American Type Culture Collection |
| Cell Strain Details: | Breast Cancer Cell lines (BT474, MCF-7, MDA-MB-231, MDA-MB-468) |
| Cell Primary Immortalized: | Immortalized |
| Species Group: | Human |
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Sample Preparation:
| Sampleprep ID: | SP000470 |
| Sampleprep Summary: | Added 1 mL of an ice-cold chloroform to the tubes and 10 ceramic beads. Tubes were vortexed for 30 seconds on a multi-tube vortexer at 5000 rpm three times to homogenize and centrifuged samples at 4 °C in a swinging bucket centrifuge for 60 min at 3,700 rpm. Majority of aqueous (top) layer was carefully transferred to 5 mL cryotubes. Majority of lipid (bottom) layer was carefully transferred to 7 mL glass vials. Remaining protein layer and residual aqueous & lipid layers were transferred into a 2 mL Lo-Bind tubes. Original tubes were rinsed with 600 µL of 2:1 chloroform:methanol solution and transferred to the 2 mL tubes. Centrifuged tubes at 15,000 rpm at 4 °C for 20 min and transferred remaining aqueous & lipid fractions into respective tubes. For each study sample, an 800 µL aliquot of the aqueous fractions was transferred to labeled 2.0 mL Lo-Bind tubes. Analytical pooled QC samples were generated by transferring an additional 75 µL aliquot from all study samples into a 10 mL tube. The total pooled sample was vortexed and 800 µL aliquots were transferred to 2.0 mL tubes labeled Pool. All tubes were frozen for 1 hr and lyophilized to dryness overnight. Samples were reconstituted by adding 700 µL of 90:10 D2O:Chenomx ISTD master mix to each, vortexed for 2 mins and centrifuged at 16,000 rcf for 4 min. A 600 µL aliquot of the supernatant was transferred into 5 mm NMR tubes (Bruker-Biospin, Switzerland), and kept on ice until data acquisition. |
| Processing Method: | lyophilization |
| Extraction Method: | Acetonitrile:water and chloroform |
| Sample Resuspension: | NMR master mix |
| Cell Type: | breast cancer |
