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MB Sample ID: SA093640
Local Sample ID: | S33 |
Subject ID: | SU001360 |
Subject Type: | Cultured cells |
Subject Species: | Mus musculus |
Taxonomy ID: | 10090 |
Select appropriate tab below to view additional metadata details:
Subject:
Subject ID: | SU001360 |
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 |
---|---|---|---|---|
S33 | SA093640 | FL013397 | Cytosol | Fraction |
S33 | SA093640 | FL013397 | Control | Treatment |
Collection:
Collection ID: | CO001354 |
Collection Summary: | Immortalized mouse macrophage-like RAW264.7 cells were obtained from the ATCC (catalog no. TIB-71). DMEM (catalog no. 10-013) and PBS (catalog no. 21-031-CV) were from Mediatech. Fetal calf serum with low endotoxin content was from Hyclone (SH30071.03 ANG19242). KLA and lipid standards were from Avanti Polar Lipids. Iodixanol (OptiPrep from Axis-Shield) was obtained from Sigma-Aldrich. Solvents were chromatography-grade and purchased from OmniSolv. All other reagents/kits were from Sigma-Aldrich. Aqueous solutions were prepared using distilled-deionized water (catalog no. 25-055-CV) from Mediatech. Isolation media were prepared K+- and Na+-free; pH was adjusted by addition of Tris base (TRIZMA). Cells were maintained, treated, and fractionated as previously described (https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2830848/). Briefly, RAW264.7 cells were maintained between passages 4 and 24 at 37°C and 10% CO2. The medium was composed of high glucose- and l-glutamine-containing DMEM supplemented with 10% heat-inactivated fetal calf serum, 100 units/ml penicillin, and 100 µg/ml streptomycin. For an experiment, five T-150 flasks of cells were plated at a density of 3.6 × 107 cells/flask in 24 ml of medium. At 24 h after plating, cells were treated with vehicle or 100 ng/ml KLA for another 24 h followed by subcellular fractionation. |
Sample Type: | Macrophages |
Treatment:
Treatment ID: | TR001375 |
Treatment Summary: | Subcellular fractionation procedures are described in detail in the publication doi: 10.1194/jlr.M008748 |
Sample Preparation:
Sampleprep ID: | SP001368 |
Sampleprep Summary: | The various extraction procedures and lipidomic analyses for each lipid class are described in the publication doi: 10.1194/jlr.M008748 |
Combined analysis:
Analysis ID | AN002141 |
---|---|
Analysis type | MS |
Chromatography type | Reversed phase/Normal phase |
Chromatography system | Several |
Column | Several |
MS Type | ESI |
MS instrument type | Several |
MS instrument name | Several |
Ion Mode | UNSPECIFIED |
Units | pmoles per mg protein |
Chromatography:
Chromatography ID: | CH001567 |
Chromatography Summary: | See publication for details doi: 10.1194/jlr.M008748 |
Instrument Name: | Several |
Column Name: | Several |
Chromatography Type: | Reversed phase/Normal phase |
MS:
MS ID: | MS001993 |
Analysis ID: | AN002141 |
Instrument Name: | Several |
Instrument Type: | Several |
MS Type: | ESI |
MS Comments: | See publication for details. doi: 10.1194/jlr.M008748 |
Ion Mode: | UNSPECIFIED |