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MB Sample ID: SA151750
| Local Sample ID: | Spender53_IL12_p |
| Subject ID: | SU001730 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | CD161+ TCR V 7.2+ MAIT cells were obtained by positive magnetic separation after isolation of Peripheral blood mononuclear cells from male human donors. |
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Subject:
| Subject ID: | SU001730 |
| Subject Type: | Cultured cells |
| Subject Species: | Homo sapiens |
| Taxonomy ID: | 9606 |
| Cell Strain Details: | CD161+ TCR V 7.2+ MAIT cells were obtained by positive magnetic separation after isolation of Peripheral blood mononuclear cells from male human donors. |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| Spender53_IL12_p | SA151750 | FL017817 | IL12/IL18 activation | Treatment |
Collection:
| Collection ID: | CO001723 |
| Collection Summary: | Peripheral blood mononuclear cells were isolated from male human donors at age 20-50 by Ficoll-Paque™ density-gradient centrifugation. CD161+ TCR Valpha 7.2+ MAIT cells were obtained by positive magnetic separation after isolation of Peripheral blood mononuclear cells from male human donors. |
| Sample Type: | Blood (whole) |
Treatment:
| Treatment ID: | TR001743 |
| Treatment Summary: | MAIT cells were stimulated with 50ng/ml IL-12 and 50ng/ml IL-18, 10µg/ml plate-bound anti-CD3 and 1µg/ml soluble anti-CD28 or a combination of both. |
Sample Preparation:
| Sampleprep ID: | SP001736 |
| Sampleprep Summary: | Cells were quenched 3 times with 1 ml ice-cold 0.9% sodium chloride. After removing the quenching solution, cells were resuspended in 100 µl ice-cold acetonitrile followed by 100 µl ice-cold Milli-Q water. After vortexing for 1 min, cells were centrifuged (14000 rpm, 4 °C, 10 min). Supernatants were transferred to new tubes. Intracellular metabolites were extracted again with 500 µl Methanol:ACN:Milli-Q water (2:3:1). After centrifugation (14000 rpm, 4 ° C, 10 min) both supernatants were combined, evaporated to dryness and stored at -80 °C until measurement. |
Combined analysis:
| Analysis ID | AN002700 |
|---|---|
| Analysis type | MS |
| Chromatography type | Reversed phase |
| Chromatography system | Agilent 1290 |
| Column | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6540 QTOF |
| Ion Mode | POSITIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH001992 |
| Instrument Name: | Agilent 1290 |
| Column Name: | Waters Acquity BEH HSS T3 (100 x 2.1mm,1.8um) |
| Column Temperature: | 45 |
| Flow Rate: | 0.3 ml/min |
| Sample Injection: | 10 µl |
| Solvent A: | 100% water; 0.1% formic acid |
| Solvent B: | 50% acetonitrile/50% MeOH; 0.1% formic acid |
| Chromatography Type: | Reversed phase |
MS:
| MS ID: | MS002498 |
| Analysis ID: | AN002700 |
| Instrument Name: | Agilent 6540 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Mass Hunter software was used to obtain raw data.Eluted metabolites were measured with the QTOF operated in centroid mode. Full scan data was generated with a scan range of 60-1600 m/z in positive ionization mode. Out of the survey scan the 5 most abundant precursor ions with charge state = 1 were subjected to fragmentation. The dynamic exclusion time was set at 30 s. Peak picking and database search was done using Progenesis QI software. |
| Ion Mode: | POSITIVE |
