Return to study ST002373 main page
MB Sample ID: SA237339
| Local Sample ID: | 10hr_M_01 |
| Subject ID: | SU002462 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 6-8 weeks |
| Gender: | Male and female |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002462 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Genotype Strain: | C57BL/6J |
| Age Or Age Range: | 6-8 weeks |
| Gender: | Male and female |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 10hr_M_01 | SA237339 | FL029390 | 10hr blank media | Treatment |
Collection:
| Collection ID: | CO002455 |
| Collection Summary: | Blank media or media post cell culture were collected and centrifuged at 1200rpm for 5min. Supernatant were flash frozen in liquid nitrogen until further analysis. |
| Sample Type: | Cultured cells |
Treatment:
| Treatment ID: | TR002474 |
| Treatment Summary: | CD8+ T cells were isolated from spleens and lymph nodes from WT mice. Cells were activated with plate-bound anti-CD3 and soluble anti-CD28. Blank media post 10, 30, 48 hours in the incubator or media post 10, 30, 48 hr activated CD8+ T cell culture were collected for MS analysis. |
Sample Preparation:
| Sampleprep ID: | SP002468 |
| Sampleprep Summary: | For measurement of extracellular metabolome, 625 µL of media was added to 375 µL acetonitrile ACN. Samples were stored at -20 °C for at least two hours followed by centrifugation at 14,000xg to precipitate any proteins. 250 µL of the supernatant was mixed with 250 µL of water and added to a 3 kDa molecular weight cut-off filter spin column (Microcon YM-3 Centrifugal Filter, Millipore). Samples were centrifuged at 14,000xg at 4 °C for 30 min. The flow-through was saved for LC-MS/MS analysis. |
Combined analysis:
| Analysis ID | AN003868 |
|---|---|
| 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: | CH002866 |
| 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: | MS003609 |
| Analysis ID: | AN003868 |
| 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 |
