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MB Sample ID: SA064554
Local Sample ID: | sgc_050516r_HL60_Zn1h_T_03 |
Subject ID: | SU001068 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Gender: | Female |
Cell Biosource Or Supplier: | ATCC |
Cell Primary Immortalized: | Yes |
Cell Passage Number: | 12 |
Cell Counts: | 1 million/mL |
Species Group: | human |
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Sample Preparation:
Sampleprep ID: | SP001075 |
Sampleprep Summary: | The remaining HL60 cells (approximately one million) were lysed in 100 µL of 1:1:2 MeCNCH3CN: CH3MeOH:NH4HCO3 (50 mM, pH 8.0) followed by one freeze-thaw cycle (three minutes each) and a ten-minute sonication in an ice bath. Protein concentration was obtained using the Pierce BCA Protein Assay Kit (Thermo Fisher Scientific). An aliquot of 100 or 150 µg of each sample in a total of 100 µL of lysis buffer was precipitated with 300 µL of ice cold 75:25 acetone:methanol for 2 h at -80°C. Samples were spun for 15 minutes at 6,800 x g and the supernatant was removed and utilized for metabolomics analysis. The pellets were rinsed with 300 µL of ice cold acetone and spun as above. Acetone was removed and the pellet was allowed to dry briefly. Metabolite supernatants were dried and reconstituted in 50 uL of appropriate reverse phase liquid chromatography (0.1% formic acid in 98:2 H2O: CH3CNMeCN) or hydrophilic interaction chromatography (80:20 CH3CNMeCN: H2O) compatible buffers prior to analyses. |