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MB Sample ID: SA237369
| Local Sample ID: | sample 1 |
| Subject ID: | SU002464 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002464 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| sample 1 | SA237369 | FL029400 | WT_Serine-free | Treatment |
Collection:
| Collection ID: | CO002457 |
| Collection Summary: | Cell pellets were spun down and washed twice with pre-warmed PBS. Metabolites were immediately extracted or stored at -80℃ until further extraction. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR002476 |
| Treatment Summary: | CD8+ T cells were isolated from spleens and lymph nodes of WT or T cell conditional GOT1 knockout mice. Cells were then activated with plate-bound anti-CD3 and soluble anti-CD28 for 24 hours in normal media (serine-replete) or serine-free media. |
Sample Preparation:
| Sampleprep ID: | SP002470 |
| Sampleprep Summary: | Cell pellets were spun down and washed twice with pre-warmed PBS. Metabolites were immediately extracted by adding methanol:water (80:20, v/v) extraction solution, sonicated and stored at -80 °C for at least 2 hours to precipitate the proteins. Supernatant after centrifugation at 14,000xg for 10 minutes was dried under nitrogen gas. Metabolites were then reconstituted using ACN:water (50:50, v/v) overnight at 4 °C. Soluble metabolites after centrifugation at 14,000xg for 10 minutes were subjected to targeted metabolite analysis by liquid-chromatography tandem mass spectrometry (LC-MS/MS). |
Combined analysis:
| Analysis ID | AN003870 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Shimadzu Prominence UFLC |
| Column | Waters XBridge BEH Amide XP HILIC (150 x 2.1mm,2.5um) |
| MS Type | ESI |
| MS instrument type | QTRAP |
| MS instrument name | ABI Sciex 5500 QTrap |
| Ion Mode | UNSPECIFIED |
| Units | AUC |
Chromatography:
| Chromatography ID: | CH002868 |
| Instrument Name: | Shimadzu Prominence UFLC |
| Column Name: | Waters XBridge BEH Amide XP HILIC (150 x 2.1mm,2.5um) |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003611 |
| Analysis ID: | AN003870 |
| Instrument Name: | ABI Sciex 5500 QTrap |
| Instrument Type: | QTRAP |
| MS Type: | ESI |
| MS Comments: | The optimized MS parameters were: ESI voltage was +5,000 V in positive ion mode and -4,500 V in negative ion mode; dwell time was 3 ms per SRM transition and the total cycle time was 1.57 seconds. Peak integration for each targeted metabolite in SRM transition was processed with MultiQuant software (v2.1, AB Sciex). The preprocessed data with integrated peak areas were exported from MultiQuant and re-imported into Metaboanalyst software for further data analysis (statistical analysis, principal component analysis, generating heatmap, enrichment analysis, etc.). |
| Ion Mode: | UNSPECIFIED |
