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MB Sample ID: SA194771
| Local Sample ID: | 0412_CD27+_Labeled_3 |
| Subject ID: | SU002153 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 6-8 weeks old |
| Weight Or Weight Range: | 15-20 grams |
| Gender: | Male and female |
| Cell Strain Details: | C57Bl/6 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU002153 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
| Age Or Age Range: | 6-8 weeks old |
| Weight Or Weight Range: | 15-20 grams |
| Gender: | Male and female |
| Cell Strain Details: | C57Bl/6 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| 0412_CD27+_Labeled_3 | SA194771 | FL023904 | CD27+ | Cell type |
Collection:
| Collection ID: | CO002146 |
| Collection Summary: | polarized CD27+ and CD27- γδ T cells were cultured in glucose-free DMEM supplemented with 10 mM [U-13C]-glucose and 10% dialyzed FBS for 24 h. The cells were rinsed in cold PBS and quenched using a mixture containing 2 mL acetonitrile and 1.5 mL H2O. After adding 1 mL chloroform, the sample was homogenized and centrifuged at 3,000 rpm, 4 oC for 20 min. The top layer was transferred to a new tube and lyophilized. The dried sample was then dissolved in 100 µL 20% acetonitrile and vigorously vortex-mixed for 3 min. After centrifugation at 14,000 rpm, 4oC for 20 min, the supernatant was collected for two-dimensional liquid chromatography-mass spectrometry (2DLC-MS) analysis. |
| Sample Type: | T-cells |
Treatment:
| Treatment ID: | TR002165 |
| Treatment Summary: | Cells were not treated with any drugs |
Sample Preparation:
| Sampleprep ID: | SP002159 |
| Sampleprep Summary: | The cells were rinsed in cold PBS and quenched using a mixture containing 2 mL acetonitrile and 1.5 mL H2O. After adding 1 mL chloroform, the sample was homogenized and centrifuged at 3,000 rpm, 4 oC for 20 min. The top layer was transferred to a new tube and lyophilized. The dried sample was then dissolved in 100 µL 20% acetonitrile and vigorously vortex-mixed for 3 min. After centrifugation at 14,000 rpm, 4oC for 20 min, the supernatant was collected for two-dimensional liquid chromatography-mass spectrometry (2DLC-MS) analysis. All samples were analyzed on a Thermo Q Exactive HF Hybrid Quadrupole-Orbitrap Mass Spectrometer coupled with a Thermo DIONEX UltiMate 3000 HPLC system (Thermo Fisher Scientific, Waltham, MA, USA). The UltiMate 3000 HPLC system was equipped with a reversed phase chromatography (RPC) column and a hydrophilic interaction chromatography (HILIC) column. The two columns were configured in parallel 2DLC mode. |
Combined analysis:
| Analysis ID | AN003376 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Q Exactive |
| Column | SeQuant ZIC-cHILIC |
| MS Type | ESI |
| MS instrument type | Orbitrap |
| MS instrument name | Thermo Exactive Plus Orbitrap |
| Ion Mode | NEGATIVE |
| Units | None |
Chromatography:
| Chromatography ID: | CH002495 |
| Chromatography Summary: | We use parallel 2DLC-MS, one column is HILIC and another is RP. |
| Instrument Name: | Q Exactive |
| Column Name: | SeQuant ZIC-cHILIC |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS003143 |
| Analysis ID: | AN003376 |
| Instrument Name: | Thermo Exactive Plus Orbitrap |
| Instrument Type: | Orbitrap |
| MS Type: | ESI |
| MS Comments: | A pooled samples for each group was detected under both positive and negative mode using 20 eV, 40eV and 60 eV. The recorded MS/MS spectrum was used for compound identification. For metabolites quantification, FullMS data of each sample was conducted under both positive and negative mode, XCMS software was used for peak deconvolution and our in-house software, MetSign was used for assignment, alignment, and statistical analysis |
| Ion Mode: | NEGATIVE |
