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MB Sample ID: SA353324
| Local Sample ID: | Serum_CreERT2_216 |
| Subject ID: | SU003357 |
| 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: | SU003357 |
| 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 |
|---|---|---|---|---|
| Serum_CreERT2_216 | SA353324 | FL041172 | serum | Sample source |
| Serum_CreERT2_216 | SA353324 | FL041172 | Gclc KO | Genotype |
Collection:
| Collection ID: | CO003350 |
| Collection Summary: | Mice were anesthetized with isoflurane, after which blood was collected via the retro-orbital venous sinus into BD microtainer tubes (BD #365967). Serum was isolated from the blood by centrifuging blood samples at 10000 xg for 5mins. Samples were stored at -80C. |
| Sample Type: | Blood (serum) |
Treatment:
| Treatment ID: | TR003366 |
| 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 serum was isolated and stored. |
Sample Preparation:
| Sampleprep ID: | SP003364 |
| Sampleprep Summary: | For serum samples, 75 µL of chloroform:methanol extraction solvent (v:v=1:2) was added to 20 µL of mouse serum. After sonicating (1400 rpm, 20°C, 5 min), the 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 | AN005303 | AN005304 |
|---|---|---|
| Analysis type | MS | MS |
| Chromatography type | Reversed phase | Reversed phase |
| Chromatography system | Thermo Vanquish | Thermo Vanquish |
| Column | Brownlee SPP C18 (75 x 2.1mm, 2.7um) | Brownlee SPP C18 (75 x 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: | CH004012 |
| Chromatography Summary: | Brownlee SPP C18 column (2.1 mm × 75 mm, 2.7 μm particle size); Solvent A is 100% H2O containing 0.1% formic acid and 1% of 1 M NH4OAc; Solvent B is 1:1 acetonitrile:isopropanol containing 0.1% formic acid and 1% of 1 M NH4OAc |
| Instrument Name: | Thermo Vanquish |
| Column Name: | Brownlee SPP C18 (75 x 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: | MS005033 |
| Analysis ID: | AN005303 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | 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: | MS005034 |
| Analysis ID: | AN005304 |
| Instrument Name: | Thermo Q Exactive HF hybrid Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | 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 |
