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MB Sample ID: SA092028
| Local Sample ID: | CB_37 |
| Subject ID: | SU001334 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Cell Strain Details: | INS-1 832/13 cell line |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001334 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Cell Strain Details: | INS-1 832/13 cell line |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| CB_37 | SA092028 | FL013200 | INS-1 (beta cells) | Cell Type |
| CB_37 | SA092028 | FL013200 | dimethylated arsenic | Treatment |
Collection:
| Collection ID: | CO001328 |
| Collection Summary: | Cells were trypsinized for approximately 30 seconds. Once collected, the cells were centrifuged at 1200 rpm for 5 minutes to form a pellet. The cell pellet was washed using PBS. |
| Sample Type: | Cultured cells |
| Storage Conditions: | -80℃ |
Treatment:
| Treatment ID: | TR001349 |
| Treatment Summary: | INS-1 832/13 cells were seeded at a density of 1 million cells per well on a 12-well plate, and exposed to 2 µM iAsIII (sodium arsenite, > 99% pure; Sigma-Aldrich, St. Louis, MO), 0.2 µM MAsIII (methylarsine oxide, > 98% pure), or 2 µM DMAsIII (Iododimethylarsine, > 98% Pure; Sigma Aldrich, St. Louis, MO) for 24h prior to glucose stimulated insulin secretion or metabolomics. Cells treated with medium without any kind of As were used as Control. |
| Cell Storage: | -80 °C |
Sample Preparation:
| Sampleprep ID: | SP001342 |
| Sampleprep Summary: | The frozen beta-cell samples were put on dry ice with a randomized order. A volume of 400 µL methanol-water (80:20) was added to the cells and vortexed by a multi-tube vortexer for 5 min at 5000 rpm. All contents in the tube were transferred into a pre-labeled MagNaLyzer tube (with 10-15 beads inside). The tubes were put on the bead homogenizer using quick run setting for bacterial cells (2 ml) with speed at 6.30 m/s for 45 sec in 1 cycle. All samples were centrifuged at 16,000 rcf for 5 min at 4°C. Supernatant from individual sample was transferred into a pre-labeled 1.5 ml low-bind Eppendorf tube. All the samples were centrifuged at 16,000 rcf for 20 min at 4°C. For quality control purpose, 32 µl of the supernatant from individual sample was transferred into a new 2-ml tube to make a sample pool (SP). Aliquots (300 µl) of supernatant from the studied samples and SP were transferred into the pre-labeled 1.5 ml Low-bind Eppendorf tubes. All samples, including studied samples and SPs, were dried using speed-vac. For immediate analysis, 100 µL of Water-Methanol (95:5) was added to the residue, and then thoroughly mixed on multiple tube vortexer for 10 mins at 5000 rpm. After centrifuge at 4°C, 16000 rcf for 4 min, the supernatant from individual sample was transferred to pre-labeled auto-sampler vial for LC-MS analysis. |
| Processing Storage Conditions: | -80℃ |
| Extract Storage: | -80℃ |
Combined analysis:
| Analysis ID | AN002103 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Thermo Vanquish |
| Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE |
| Units | Relative Intensity |
Chromatography:
| Chromatography ID: | CH001535 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| Column Pressure: | 6000-10000 psi |
| Column Temperature: | 50 |
| Flow Rate: | 0.4 ml/min |
| Injection Temperature: | 8 |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 100% methanol; 0.1% formic acid |
| Analytical Time: | 22 min |
| Capillary Voltage: | 3.75 KV |
| Weak Wash Solvent Name: | 10:90 Methanol:Water with 0.1% FA solution |
| Strong Wash Solvent Name: | 75:25 2-Propanol: Water with 0.1% FA solution |
| Randomization Order: | Yes |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS001954 |
| Analysis ID: | AN002103 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | We used DDA mode to acquire the MS and MS/MS data. Progenesis QI was used for peak picking, alignment, and normalization. |
| Ion Mode: | POSITIVE |
| Capillary Temperature: | 275 °C |
| Capillary Voltage: | 3.75 KV |
| Collision Energy: | 10-35, ramp |
| Collision Gas: | N2 |
| Dry Gas Flow: | 45 |
| Dry Gas Temp: | 325°C |
| Fragmentation Method: | CID |
| Ionization: | ES+ |
| Mass Accuracy: | 5ppm |
| Dataformat: | Profile |
| Desolvation Gas Flow: | 45 |
| Desolvation Temperature: | 325°C |
| Resolution Setting: | 120000 |
| Scan Range Moverz: | 70-1050 m/z |
| Scanning Range: | 70-1050 m/z |
