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MB Sample ID: SA093650
| Local Sample ID: | S15 |
| Subject ID: | SU001360 |
| Subject Type: | Cultured cells |
| Subject Species: | Mus musculus |
| Taxonomy ID: | 10090 |
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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 |
