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MB Sample ID: SA165052
Local Sample ID: | 6h ATP_2 |
Subject ID: | SU001856 |
Subject Type: | Cultured cells |
Subject Species: | Homo sapiens |
Taxonomy ID: | 9606 |
Cell Biosource Or Supplier: | ATCC |
Cell Strain Details: | Human Lung epithelial A549 cancer cells |
Cell Passage Number: | 10 |
Cell Counts: | 5 million cells |
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Sample Preparation:
Sampleprep ID: | SP001862 |
Sampleprep Summary: | 5 × 106 A549 cells were treated with control (no treatment), 0.5 mM ATP or 10 ng/ml TGF-beta for 2, 6 and 12 hours. After treatment, cells were washed twice with deionized water and polar metabolites were then extracted with cryogenically cold 80% methanol/water mixture. LC-MS grade water, methanol, and acetonitrile (Fischer Scientific, PA, USA) were used. Methanol-extracted samples were then sonicated in cycles of sonication phase and rest phase for 10 minutes (5 second sonication phase and 10 seconds halt). The samples were then centrifuged at 13,000 rpm for 10 minutes and supernatant was then collected. Supernatants collected from in vitro and in vivo extraction were then lyophilized. Briefly, the supernatant was then lyophilized by using a speed vacuum evaporator. The samples were then dissolved into a mixture of acetonitrile/water (1:1; v/v). |
Sampleprep Protocol Filename: | Sample preparation protocol |
Processing Method: | CEll scrapping Quenching |
Processing Storage Conditions: | -80℃ |
Extraction Method: | Quenching with Ice cold methanol |
Extract Enrichment: | Speed vaccum evaporator |
Extract Storage: | -80℃ |
Sample Resuspension: | Acetonitrile/water (1:1) |