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MB Sample ID: SA163298
| Local Sample ID: | LPS13C3 |
| Subject ID: | SU001821 |
| Subject Type: | Cultured cells |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Select appropriate tab below to view additional metadata details:
Subject:
| Subject ID: | SU001821 |
| Subject Type: | Cultured cells |
| Subject Species: | Rattus norvegicus |
| Taxonomy ID: | 10116 |
Factors:
| Local Sample ID | MB Sample ID | Factor Level ID | Level Value | Factor Name |
|---|---|---|---|---|
| LPS13C3 | SA163298 | FL019445 | 1 | LPS (ug/mL) |
| LPS13C3 | SA163298 | FL019445 | U-13C Glucose | Glucose |
Collection:
| Collection ID: | CO001814 |
| Collection Summary: | Immortalized rat astrocytes (CRL-2005) |
| Sample Type: | Fibroblasts |
Treatment:
| Treatment ID: | TR001834 |
| Treatment Summary: | high-glucose DMEM (Gibco) containing 10% FBS and 1% penicillin/streptomycin (Gibco). Isotopic labeling was achieved through 30 min treatments with media containing 4.5 g/L U-13C-glucose (100% of the total glucose content; Cambridge Isotopes); parallel cultures were treated with 4.5 g/L natural-abundance glucose. Simultaneous to the introduction of the labeled substrate, subsets of both unlabeled and labeled cultures were treated with LPS (added to culture media to a final concentration of 1 μg/mL) for the duration of the labeling protocol. |
Sample Preparation:
| Sampleprep ID: | SP001827 |
| Sampleprep Summary: | Cells were then washed with phosphate buffer solution and high-performance liquid chromatography (HPLC)-grade water, quenched with 1 mL cold HPLC-grade methanol, scraped from the plate, and pelleted. Pellets were dried on a SpeedVac and subsequently lyophilized. Dried samples were weighed out and extracted with the solvent volumes adjusted to maintain a ratio of 1 mL of solvent per 1 mg of dried cellular material. The final volume of reconstitution solvent was adjusted to 100 μL per 1 mg of dried material. All cell-culture conditions (unlabeled versus labeled, control versus LPSstimulated) were sampled in triplicate. |
Combined analysis:
| Analysis ID | AN002837 |
|---|---|
| Analysis type | MS |
| Chromatography type | HILIC |
| Chromatography system | Agilent 1200 |
| Column | Luna Aminopropyl |
| MS Type | ESI |
| MS instrument type | QTOF |
| MS instrument name | Agilent 6520 QTOF |
| Ion Mode | NEGATIVE |
| Units | peak area |
Chromatography:
| Chromatography ID: | CH002100 |
| Instrument Name: | Agilent 1200 |
| Column Name: | Luna Aminopropyl |
| Chromatography Type: | HILIC |
MS:
| MS ID: | MS002630 |
| Analysis ID: | AN002837 |
| Instrument Name: | Agilent 6520 QTOF |
| Instrument Type: | QTOF |
| MS Type: | ESI |
| MS Comments: | Mass spectrometry (MS) detection was carried out on an Agilent 6520 Q-TOF in negative ESI (electrospray ionization) mode with the following settings: gas temperature 325 °C, drying gas 5 L/min, nebulizer 15 psi, fragmentor 120 V, skimmer 65 V, capillary voltage −3500 V, and scan rate 1.06 spectra/s. Tandem MS (MS/MS) analyses were carried out with identical ESI parameters, and the following fragmentation and precursor ion selection settings: collision energy 10 V, precursor isolation window 1.3 amu, and scan rate 1.00 spectra/s. X13CMS R-Package was used for data processing |
| Ion Mode: | NEGATIVE |
