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MB Sample ID: SA064613
| Local Sample ID: | sds_20160607s_48h_HILIC_Zn_HL60_challenge_Con2 |
| 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 |
Select appropriate tab below to view additional metadata details:
Subject:
| 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 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| sds_20160607s_48h_HILIC_Zn_HL60_challenge_Con2 | SA064613 | FL011022 | 48 hr | time point |
| sds_20160607s_48h_HILIC_Zn_HL60_challenge_Con2 | SA064613 | FL011022 | Control | sample type |
Collection:
| Collection ID: | CO001062 |
| Collection Summary: | no collection protocol |
| Sample Type: | Leukemia cells |
Treatment:
| Treatment ID: | TR001082 |
| Treatment Summary: | The HL60 cells were cultured in Isocove’s Modified Dulbecco’s Medium (IMDM, Gibco) supplemented with 10% v/v heat-inactivated fetal bovine serum (Atlanta Biologicals) at 37˚C with 5% CO2 atmosphere and treated with 225 µM Zn or deionized H2O. |
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. |
Combined analysis:
| Analysis ID | AN001687 | AN001688 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | HILIC |
| Chromatography system | Waters NanoAcquity | Waters Acquity |
| Column | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) | Phenomenex Kinetex HILIC 100A (50 x 2.1 mm,2.6um) |
| MS Type | ESI | ESI |
| MS instrument type | QTOF | QTOF |
| MS instrument name | Waters Synapt G2 S QTOF | Waters Synapt G2 S QTOF |
| Ion Mode | POSITIVE | POSITIVE |
| Units | absorbance | Abundance |
Chromatography:
| Chromatography ID: | CH001187 |
| Chromatography Summary: | see formatted metadata template |
| Instrument Name: | Waters NanoAcquity |
| Column Name: | Waters Acquity HSS T3 (100 x 2.1mm,1.8um) |
| Chromatography Type: | Reversed phase |
| Chromatography ID: | CH001188 |
| Chromatography Summary: | see formatted metadata template |
| Instrument Name: | Waters Acquity |
| Column Name: | Phenomenex Kinetex HILIC 100A (50 x 2.1 mm,2.6um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS001562 |
| Analysis ID: | AN001687 |
| Instrument Name: | Waters Synapt G2 S QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
| MS ID: | MS001563 |
| Analysis ID: | AN001688 |
| Instrument Name: | Waters Synapt G2 S QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| Ion Mode: | POSITIVE |
