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MB Sample ID: SA353359
| Local Sample ID: | Kidney_FF_225 |
| Subject ID: | SU003361 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU003361 |
| Subject Type: | Mammal |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Kidney_FF_225 | SA353359 | FL041181 | kidney | Sample source |
| Kidney_FF_225 | SA353359 | FL041181 | Gclc WT | Genotype |
Collection:
| Collection ID: | CO003354 |
| Collection Summary: | Mice were euthanized with CO2 inhalation. Tissues were isolated, snap frozen on dry ice, and stored at -80C. |
| Sample Type: | Kidney |
Treatment:
| Treatment ID: | TR003370 |
| Treatment Summary: | Gclc WT (Gclc f/f) and whole-body Gclc KO (Gclc f/f Rosa26-CreERT2) were treated with 160 mg/kg i.p. injection of tamoxifen daily for five consecutive days. After approximately 14 days, mice were euthanized and tissue was isolated and stored. |
Sample Preparation:
| Sampleprep ID: | SP003368 |
| Sampleprep Summary: | Tissues were homogenized with a pre-chilled BioPulverizer (59012MS, BioSpec) and then placed on dry ice. The chloroform:methanol extraction solvent (v:v = 1:1) was added to the homogenate to meet 50 mg/mL. The samples were then sonicated in ice-cold water using BiorupterTM UCD-200 sonicator for 5 min (30 s sonication and 30 s rest cycle; high voltage mode). The lipid extracts were cleared by centrifugation (17,000g, 20°C, 10 min), and the lipids in the supernatant were analyzed by LC-MS. |
Combined analysis:
| Analysis ID | AN005311 | AN005312 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um) | Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um) |
| MS Type | ESI | ESI |
| MS instrument type | Orbitrap | Orbitrap |
| MS instrument name | Thermo Q Exactive HF hybrid Orbitrap | Thermo Q Exactive HF hybrid Orbitrap |
| Ion Mode | POSITIVE | NEGATIVE |
| Units | Normalized to the median value of total lipid signals | Normalized to the median value of total lipid signals |
Chromatography:
| Chromatography ID: | CH004016 |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Perkin Elmer Brownlee SPP C18 (75 × 2.1mm, 2.7um) |
| Column Temperature: | 50 |
| Flow Gradient: | 0–2 min 35% B, 2–8 min from 35 to 80% B, 8–22 min from 80 to 99% B, 22–36 min 99% B, and 36.1–40 min from 99 to 35% B. |
| Flow Rate: | 0.400 mL/min |
| Solvent A: | 100% water; 0.1% formic acid; 10mM ammonium acetate |
| Solvent B: | 50% acetonitrile/50% isopropanol; 0.1% formic acid; 10mM ammonium acetate |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS005041 |
| Analysis ID: | AN005311 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | For the mass spectrometry, the data-dependent MS2 scan conditions were applied in both positive and negative mode: the scan range was from m/z 250–1500, resolution was 60,000 for MS, and 30,000 for DDMS2 (top 10), and the AGC target was 3E6 for full MS and 1E5 for DDMS2, allowing ions to accumulate for up to 200 ms for MS and 50 ms for MS/MS. For MS/MS, the following settings are used: isolation window width 1.2 m/z with an offset of 0.5 m/z, stepped NCE at 10, 15, and 25 a.u., minimum AGC 5E2, and dynamic exclusion of previously sampled peaks for 8 s. |
| Ion Mode: | POSITIVE |
| MS ID: | MS005042 |
| Analysis ID: | AN005312 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | For the mass spectrometry, the data-dependent MS2 scan conditions were applied in both positive and negative mode: the scan range was from m/z 250–1500, resolution was 60,000 for MS, and 30,000 for DDMS2 (top 10), and the AGC target was 3E6 for full MS and 1E5 for DDMS2, allowing ions to accumulate for up to 200 ms for MS and 50 ms for MS/MS. For MS/MS, the following settings are used: isolation window width 1.2 m/z with an offset of 0.5 m/z, stepped NCE at 10, 15, and 25 a.u., minimum AGC 5E2, and dynamic exclusion of previously sampled peaks for 8 s. |
| Ion Mode: | NEGATIVE |
