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MB Sample ID: SA218462

Local Sample ID:WT3
Subject ID:SU002366
Subject Type:Mammal
Subject Species:Mus musculus
Taxonomy ID:10090

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Sample Preparation:

Sampleprep ID:SP002372
Sampleprep Summary:One million CD45+ F4/80+ CD11c+ FACS-sorted alveolar macrophages (AMs) from the bronchoalveolar lavage (BAL) of adult Tfamf/f and CD11c∆Tfam mice were collected. Each sample was generated by merging FACS-sorted AMs from 13 to 30 mice. Quality Control (QC) samples (n=4) were prepared by pooling equal volumes of cell extracts from each sample by following the same protocols used for the subject samples. Samples were subjected to two freeze–thaw cycles for metabolism quenching and complete metabolite extraction, specifically by placing the samples at -80ºC for 15 min and thawing them on ice for 10 min with brief vortex-mixing. The samples were then centrifuged at 20,000 xg at 4°C for 10 min and the supernatant collected. The supernatant was evaporated to dryness (SpeedVac Concentrator, Thermo Fisher Scientific, Waltham, MA, USA) and derivatized with 10 μl O-methoxyamine hydrochloride (15mg/mL) in pyridine and 10 μl N,O-bis(trimethylsilyl)trifluoroacetamide in 1% trimethylchlorosilane. Finally, 100 μl of heptane containing 10 ppm of 4-nitrobenzoic acid (IS) was used as internal standard to monitor sample injection
Sampleprep Protocol Filename:Protocol_AM.pdf