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MB Sample ID: SA081604
Local Sample ID: | FOXO1-ER1 |
Subject ID: | SU001244 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
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Sample Preparation:
Sampleprep ID: | SP001252 |
Sampleprep Summary: | The cells treated with vehicle (control, N = 3) or tamoxifen (FOXO1-ER, N = 3) were used for the metabolomic analysis (Human Metabolome Technologies Inc., Tsuruoka, Japan). 3.9 x 106 cells were transferred into 500 μL of methanol containing 50 mM external standard. After homogenization by BMSM10N21 (BMS, Tokyo) at 1,500 rpm for 120 s performed 5 times, 500 μL of chloroform and 200 μL of ultrapure water were added to the homogenate. The solution was mixed well and centrifuged at 2,300 × g for 5 min at 4 °C. The resultant water phase was ultrafiltered using a Millipore Ultrafree-MC PLHCC HMT Centrifugal Filter Device, 5 kDa (Millipore, Billerica, MA). The filtrate was dried and then dissolved in 50 μL of ultrapure water. The samples obtained were then subjected to capillary electrophoresis time-of-flight mass spectrometry (CE-TOFMS) analysis using the Agilent CE-TOFMS system (Agilent Technologies, Santa Clara, CA) at 4 °C. The detected peaks were aligned according to their m/z values and normalized migration times. The peaks were mean-centered and scaled using their standard deviations on a per peak basis as a pretreatment. |