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MB Sample ID: SA085829
Local Sample ID: | 4661 02 |
Subject ID: | SU001280 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001280 |
Subject Type: | Mammal |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Gender: | Male |
Factors:
Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
---|---|---|---|---|
4661 02 | SA085829 | FL012835 | 5C | Temperature |
4661 02 | SA085829 | FL012835 | WT | Genotype |
Collection:
Collection ID: | CO001274 |
Collection Summary: | Mice were anaesthetised with Isoflurane (under controlled flow-rate), and placed in a hot pad, while the blood was drawn from the tail into a tube. The blood was left at 22C for 30 - 60 minutes, and then centrifuged at 14,000 RPM for 3 minutes. The serum fraction was collected and frost at -20C. |
Sample Type: | Blood (serum) |
Treatment:
Treatment ID: | TR001295 |
Treatment Summary: | Wild-type and Ucp1CRE/12-LOX KO mice were exposed to a short-term cold temperature (1 hour at 5C) or kept at room temperature. After this period, the serum was collected as described. Ucp1CRE/12-LOX KO mice were created through CRISPR-Cas9 technology, as described in Leiria et al., 2019, Cell Metabolism. |
Sample Preparation:
Sampleprep ID: | SP001288 |
Sampleprep Summary: | Aliquots of 100 µL serum or 1mg protein from homogenized tissue (measured by BCA) were taken, depending on the experiment. A mixture of deuterium-labeled internal standards was added to each aliquot, followed by 3x volume of sample of cold methanol (MeOH). Samples were vortexed for 5 minutes and stored at 31 −20 °C overnight. Cold samples were centrifuged at 14,000g for 10 minutes, and the supernatant was then transferred to a new tube and 3 mL of acidified H 2 O (pH 3.5) was added to each sample prior to C18 SPE. The methyl formate fractions were collected, dried under nitrogen, and reconstituted in 50 µL MeOH:H 2 O (1:1, by vol). Samples were transferred to 0.5 mL tubes and centrifuged at 20,000g at 4 °C for 10 minutes. 35ul of supernatant was transferred to LC–MS/MS vials for analysis using the BERG LCMS/MS mediator lipidomics platform. |
Combined analysis:
Analysis ID | AN002024 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase |
Chromatography system | Ekspert MicroLC 200 system |
Column | Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um) |
MS Type | ESI |
MS instrument type | Triple TOF |
MS instrument name | ABI Sciex 5600+ TripleTOF |
Ion Mode | NEGATIVE |
Units | Peak Area |
Chromatography:
Chromatography ID: | CH001465 |
Instrument Name: | Ekspert MicroLC 200 system |
Column Name: | Phenomenex Synergi Fusion-RP capillary C18 (150 × 0.5 mm,4um) |
Chromatography Type: | Reversed phase |
MS:
MS ID: | MS001877 |
Analysis ID: | AN002024 |
Instrument Name: | ABI Sciex 5600+ TripleTOF |
Instrument Type: | Triple TOF |
MS Type: | ESI |
MS Comments: | MS spectra were acquired in high-resolution mode (>30,000) using a 100-ms accumulation time per spectrum. Fullscan MS/MS was acquired in high sensitivity mode, with an accumulation time optimized per cycle. Collision energy was set using rolling collision energy with a spread of 15V. The identity of a component was confirmed using PeakView® software (SCIEX), and quantification was performed using MultiQuant™ software (SCIEX). |
Ion Mode: | NEGATIVE |