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Metabolomics raw data and table of extracted peak areas for BT478, BT530 and NHA cells cultured in the presence of MPA, Compound 3 (IC80) or vehicle control, for 6 hours.

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DataCite Commons2024-10-08 更新2025-01-06 收录
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https://figshare.com/articles/dataset/Metabolomics_raw_data_and_table_of_extracted_peak_areas_for_BT478_BT530_and_NHA_cells_cultured_in_the_presence_of_MPA_Compound_3_IC80_or_vehicle_control_for_6_hours_/22246258
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Sample Collection For metabolomics profiling, 10^6 cells (BT478, BT530, NHAs) were cultured in the presence of MPA, Compound 3 (IC80) or vehicle control, for 6 hours, and then collected, washed with PBS, and flash frozen in liquid nitrogen.  Sample Extraction Metabolites were extracted with a solution of cold acetonitrile/methanol/water (2:2:1) from the cell pellets and protein precipitation was performed by three cycles of freeze/thawing and sonication.  Sample analysis The LC-MS metabolomics analysis was performed using a UHPLC-MS system consisting of an Agilent 6550 qToF coupled to an Agilent 1290 binary pump UHPLC system. The source parameters were as follows: Gas temperature, 150°C at 14 L/min and 45 psig; Sheath gas temperature, 325°C at 12 L/min; Capillary and nozzle voltages were set to -2.0 kV. iFunnel conditions were changed from default to-30 V DC, High pressure funnel drop -100 V and RF voltage of 110 V, low pressure funnel drop -50 V and RF voltage of 60 V. Chromatographic separation was achieved by ion-paired chromatography. In brief, 2 mL of each sample was injected onto Agilent ZORBAX Extend-C18 (150 mm 3 2.1 mm i.d.; 1.8 mm) column using tributylamine (TBA) as an ion paring agent (solvent A: 3% methanol, 97% water 10 mM TBA, 15 mM Acetic acid, solvent B: 100% methanol). The linear gradient employed was as follows: 0–2.5 min 99% A, 2.5–7.5 min decrease to 80% A, 7.5–13 min to 55% B 652 and finally 13–15 min to 99% B and held for 1 min. The column was re- generated for 2 min at 1% B. The flow rate was set to 250 mL/min. The column temperature was maintained at 25°C.  Data processing Skyline was used for data processing of metabolites in our library of standards using known retention times and MS/MS spectra. Integrated peak areas for the metabolites was exported for further statistical and metabolite enrichment analysis by using online MetaboAnalyst (https://www.metaboanalyst.ca/).
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figshare
创建时间:
2023-09-25
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